Cold menthol receptor antagonists

ABSTRACT

Disclosed are compounds, compositions and methods for treating various diseases, syndromes, conditions and disorders, including pain. Such compounds are represented by Formula (I) as follows: 
                         
wherein A, L, R 1 , R 2 , R 4  and R 5  are defined herein.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Patent ApplicationNo. 61/089,933, filed Aug. 19, 2008, currently pending, which is herebyincorporated by reference in its entirety.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

The research and development of the invention described below was notfederally sponsored.

BACKGROUND OF THE INVENTION

Transient receptor potential (TRP) channels are non-selective cationchannels that are activated by a variety of stimuli. Numerous members ofthe ion channel family have been identified to date, including thecold-menthol receptor, also called TRPM8 (McKemy D. D., et al., Nature2002, 416(6876), 52-58). Collectively, the TRP channels and relatedTRP-like receptors connote sensory responsivity to the entire continuumof thermal exposure, selectively responding to threshold temperaturesranging from noxious hot through noxious cold as well as to certainchemicals that mimic these sensations. Specifically, TRPM8 is known tobe stimulated by cool to cold temperatures as well as by chemicalagents, such as, menthol and icilin, which may be responsible for thetherapeutic cooling sensation that these agents provoke.

TRPM8 is located on primary nociceptive neurons (A-delta and C-fibers)and is also modulated by inflammation-mediated second messenger signals(Abe, J., et al., Neurosci Lett 2006, 397(1-2), 140-144; Premkumar, L.S., et al., J. Neurosci, 2005, 25(49), 11322-11329). The localization ofTRPM8 on both A-delta and C-fibers may provide a basis for abnormal coldsensitivity in pathologic conditions wherein these neurons are altered,resulting in pain, often of a burning nature (Kobayashi, K., et al., JComp Neurol, 2005, 493(4), 596-606; Roza, C., et al., Pain, 2006,120(1-2), 24-35; and Xing, H., et al., J Neurophysiol, 2006, 95(2),1221-30). Cold intolerance and paradoxical burning sensations induced bychemical or thermal cooling closely parallel symptoms seen in a widerange of clinical disorders and thus provide a strong rationale for thedevelopment of TRPM8 modulators as novel antihyperalgesic orantiallodynic agents. TRPM8 is also known to be expressed in the brain,lung, bladder, gastrointestinal tract, blood vessels, prostate andimmune cells, thereby providing the possibility for therapeuticmodulation in a wide range of maladies.

International patent application WO 2006/040136 A1 from Bayer HealthcareAG purportedly describes substituted 4-benzyloxy-phenylmethylamidederivatives as cold menthol receptor-1 (CMR-1) antagonists for thetreatment of urological disorders. International patent application WO2006/040103 A1, from Bayer Healthcare AG purportedly describes methodsand pharmaceutical compositions for treatment and/or prophylaxis ofrespiratory diseases or disorders.

International patent applications WO 2007/017092A1, WO 2007/017093A1 andWO 2007/017094A1, from Bayer Healthcare AG, purportedly describebenzyloxyphenylmethyl carbamate, substituted 2-benzyloxybenzoic acidamide and substituted 4-benzyloxybenzoic acid amide derivatives for thetreatment of diseases associated with the cold menthol receptor (CMR),a.k.a. TRPM8.

There is a need in the art for TRPM8 antagonists that can be used totreat a disease, syndrome, or condition in a mammal in which thedisease, syndrome or condition is affected by the modulation of TRPM8receptors, such as pain, the diseases that lead to such pain, andpulmonary or vascular dysfunction.

SUMMARY OF THE INVENTION

The present invention is directed to compounds of Formula (I)

wherein

-   A is phenyl, thienyl, or C₅₋₇cycloalkyl;-   provided that when A is phenyl, R₂ is other than 2-chloro,    2-C₁₋₄alkoxy, 4-C₁₋₄alkoxy, 2-C₁₋₄alkoxycarbonyl, or    4-C₁₋₄alkoxycarbonyl;-   or, when A is phenyl, L is —N(R₃)(CH₂)_(n)—, and R₁ and L are    attached to adjacent carbon atoms of phenyl, R₁ and R₃ are    optionally taken with the atoms to which they are attached to form    dihydro-indol-1-yl or dihydro-quinolin-1-yl;-   R₁ is one to three substituents selected from the group consisting    of hydrogen, C₁₋₄alkyl, C₁₋₄alkoxy, fluoro, chloro, and hydroxy;    -   L is -Z-(CH₂)_(n)— or —CH₂—;    -   n is 0 or 1;-   and Z is O, S, or NR₃; provided that when Z is O or S, A is phenyl;-   R₂ is hydrogen, C₁₋₄alkyl, C₁₋₄alkoxy, trifluoromethyl, chloro,    fluoro, bromo, carboxy, or C₁₋₄alkoxycarbonyl;-   R₃ is hydrogen or C₁₋₃alkyl;-   R₄ is hydrogen or C₁₋₄alkyl;-   R₅ is naphthyl, indanyl, tetralinyl, or a 9 to 10-membered    heteroaryl selected from the group consisting of benzimidazolyl,    cinnolinyl, 1H-indazolyl, indolyl, isoindolyl, isoquinolinyl,    naphthyridinyl, phthalazinyl, quinazolinyl, quinolinyl, and    quinoxalinyl; wherein R₅ is optionally substituted with 1 to 2    substituents independently selected from the group consisting of    C₁₋₄alkyl, hydroxy, C₁₋₄alkoxy, trifluoromethyl, chloro, fluoro,    bromo, carboxy, C₁₋₄alkoxycarbonyl, and cyano;-   and enantiomers, diastereomers, solvates and pharmaceutically    acceptable salts thereof;-   provided that a compound of Formula (I) is other than-   a compound wherein A is phenyl, R₁ is 3-chloro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, n is 1, Z is NR₃, R₃ is    hydrogen, R₄ is hydrogen, and R₅ is isoquinolin-5-yl;-   a compound wherein A is phenyl, R₁ is 2-fluoro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ is    hydrogen, R₄ is hydrogen, and R₅ is indol-4-yl;-   and-   a compound wherein A is phenyl, R₁ is 4-methoxy, R₂ is 4-chloro, L    is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ is hydrogen, R₄ is hydrogen,    and R₅ is isoquinolin-5-yl.

Illustrative of the invention is a pharmaceutical compositioncomprising, consisting of and/or consisting essentially of apharmaceutically acceptable carrier, a pharmaceutically acceptableexcipient, and/or a pharmaceutically acceptable diluent and a compoundof Formula (I). Also illustrative of the invention is a process formaking a pharmaceutical composition comprising, consisting of, and/orconsisting essentially of admixing a compound of Formula (I) and apharmaceutically acceptable carrier, a pharmaceutically acceptableexcipient, and/or a pharmaceutically acceptable diluent.

The present invention is further directed to methods for treating orameliorating a disease or condition in a subject, including a mammal,and/or a human in which the disease or condition is affected by themodulation of TRPM8 receptors, such as pain, the diseases that lead tosuch pain, and pulmonary or vascular dysfunction using a compound ofFormula (I). In particular, the method of the present invention isdirected to treating or ameliorating a TRPM8 receptor-modulateddisorders including, inflammatory pain, cold-intolerance or coldallodynia, peripheral vascular pain, itch, urinary incontinence, chronicobstructive pulmonary disease, pulmonary hypertension, and anxiety,including other stress-related disorders using a compound of Formula(I).

The present invention is also directed to methods for producing theinstant compounds and pharmaceutical compositions and medicamentsthereof.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, with reference to substituents, the term “independently”means that when more than one of such substituent is possible, suchsubstituents may be the same or different from each other.

As used herein, unless otherwise noted, “alkyl” whether used alone or aspart of a substituent group refers to straight and branched carbonchains having 1 to 8 carbon atoms or any number within this range.Therefore, designated numbers of carbon atoms (e.g. C₁₋₈) shall referindependently to the number of carbon atoms in an alkyl moiety or to thealkyl portion of a larger alkyl-containing substituent. In substituentgroups with multiple alkyl groups, such as, (C₁₋₆alkyl)₂amino- theC₁₋₆alkyl groups of the dialkylamino may be the same or different.

As used herein, unless otherwise noted, the term “alkoxy” refers to an—Oalkyl substituent group, wherein alkyl is as defined supra. To theextent substituted, an alkyl and alkoxy chain may be substituted on acarbon atom.

As used herein, the terms “alkenyl” and “alkynyl” refer to straight andbranched carbon chains having 2 or more carbon atoms, wherein an alkenylchain has at least one double bond in the chain and an alkynyl chain hasat least one triple bond in the chain.

As used herein, the term “cycloalkyl” refers to saturated or partiallyunsaturated, monocyclic or polycyclic hydrocarbon rings of from 3 to 14carbon atom members. Examples of such rings include, and are not limitedto cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl andadamantyl. Similarly, “cycloalkenyl” refers to a cycloalkyl thatcontains at least one double bond in the ring. Additionally, a“benzo-fused cycloalkyl” is a cycloalkyl ring that is fused to a benzenering. A “heteroaryl-fused cycloalkyl” is a cycloalkyl ring that is fusedto a 5 or 6 membered heteroaryl ring (containing one of O, S or N and,optionally, one additional nitrogen).

As used herein, the term “heterocyclyl” refers to a nonaromatic cyclicring of 5 to 7 members in which 1 to 2 members are nitrogen, or anonaromatic cyclic ring of 5 to 7 members in which zero, one or twomembers are nitrogen and up to two members are oxygen or sulfur;wherein, optionally, the ring contains zero to one unsaturated bonds,and, optionally, when the ring is of 6 or 7 members, it contains up totwo unsaturated bonds. The term “benzo-fused heterocyclyl” includes a 5to 7 membered monocyclic heterocyclic ring fused to a benzene ring. Theterm “heteroaryl-fused heterocyclyl” refers to 5 to 7 memberedmonocyclic heterocyclic ring fused to a 5 or 6 membered heteroaryl ring(containing one of O, S or N and, optionally, one additional nitrogen).The term “cycloalkyl-fused heterocyclyl” refers to a 5 to 7 memberedmonocyclic heterocyclic ring fused to a 5 to 7 membered cycloalkyl orcycloalkenyl ring. Furthermore, the term “heterocyclyl-fusedheterocycyl” refers to a 5 to 7 membered monocyclic heterocyclic ringfused to a 5 to 7 membered heterocyclyl ring (of the same definition asabove but absent the option of a further fused ring). For instantcompounds of the invention, the carbon atom ring members that form theheterocyclyl ring are fully saturated. Other compounds of the inventionmay have a partially saturated heterocyclyl ring. The term“heterocyclyl” also includes a 5 to 7 membered monocyclic heterocyclebridged to form bicyclic rings. Such compounds are not considered to befully aromatic and are not referred to as heteroaryl compounds. Examplesof heterocyclyl groups include, and are not limited to, pyrrolinyl(including 2H-pyrrole, 2-pyrrolinyl or 3-pyrrolinyl), pyrrolidinyl,2-imidazolinyl, imidazolidinyl, 2-pyrazolinyl, pyrazolidinyl,piperidinyl, morpholinyl, thiomorpholinyl and piperazinyl.

As used herein, the term “aryl” refers to an unsaturated, aromaticmonocyclic ring of 6 carbon members or to an unsaturated, aromaticpolycyclic ring of from 10 to 14 carbon members. Examples of such arylrings include, and are not limited to, phenyl, naphthalenyl oranthracenyl. Preferred aryl groups for the practice of this inventionare phenyl and naphthalenyl.

As used herein, the term “heteroaryl” refers to an aromatic ring of 5 or6 members wherein the ring consists of carbon atoms and has at least oneheteroatom member. Suitable heteroatoms include nitrogen, oxygen orsulfur. In the case of 5 membered rings, the heteroaryl ring containsone member of nitrogen, oxygen or sulfur and, in addition, may containup to three additional nitrogens. In the case of 6 membered rings, theheteroaryl ring may contain from one to three nitrogen atoms. For thecase wherein the 6 membered ring has three nitrogens, at most twonitrogen atoms are adjacent.

Optionally, the heteroaryl ring is fused to a benzene ring to form a“benzo fused heteroaryl”; similarly, the heteroaryl ring is optionallyfused to a 5 or 6 membered heteroaryl ring (containing one of O, S or Nand, optionally, one additional nitrogen) to form a “heteroaryl-fusedheteroaryl”; similarly, the heteroaryl ring is optionally fused to a 5to 7 membered cycloalkyl ring or a 5 to 7 membered heterocyclo ring (asdefined supra but absent the option of a further fused ring) to form a“cycloalkyl-fused heteroaryl”. Examples of heteroaryl groups include,and are not limited to, furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl,imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl,thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl or pyrazinyl; examplesof heteroaryl groups with the optionally fused benzene rings includeindolyl, isoindolyl, indolinyl, benzofuryl, benzothienyl, indazolyl,benzimidazolyl, benzthiazolyl, benzoxazolyl, benzisoxazolyl,benzothiadiazolyl, benzotriazolyl, quinolizinyl, quinolinyl,isoquinolinyl or quinazolinyl.

As used herein, the term “arylalkyl” means an alkyl group substitutedwith an aryl group (e.g., benzyl, phenethyl). Similarly, the term“arylalkoxy” indicates an alkoxy group substituted with an aryl group(e.g., benzyloxy).

As used herein, the term “halogen” refers to fluorine, chlorine, bromineand iodine. Substituents that are substituted with multiple halogens aresubstituted in a manner that provides compounds that are stable.

As used herein, the term “R” at a stereocenter designates that thestereocenter is purely of the R-configuration as defined in the art;likewise, the term “S” means that the stereocenter is purely of theS-configuration. As used herein, the terms “*R” or “*S” at astereocenter are used to designate that the stereocenter is of pure butunknown configuration. As used herein, the term “RS” refers to astereocenter that exists as a mixture of the R- and S-configurations.

Compounds containing one stereocenter drawn without a stereo bonddesignation are a mixture of two enantiomers. Thus, stereocenters drawnwithout stereo bond designations are a mixture of R- andS-configurations. For stereocenters drawn with stereo bond designations,the absolute stereochemistry is as depicted.

Whenever the term “alkyl” or “aryl” or either of their prefix rootsappear in a name of a substituent (e.g., arylalkyl, alkylamino) the nameis to be interpreted as including those limitations given above for“alkyl” and “aryl.” Designated numbers of carbon atoms (e.g., C₁-C₆)refer independently to the number of carbon atoms in an alkyl moiety, anaryl moiety, or in the alkyl portion of a larger substituent in whichalkyl appears as its prefix root. For alkyl and alkoxy substituents, thedesignated number of carbon atoms includes all of the independentmembers included within a given range specified. For example C₁₋₆ alkylwould include methyl, ethyl, propyl, butyl, pentyl and hexylindividually as well as sub-combinations thereof (e.g. C₁₋₂, C₁₋₃, C₁₋₄,C₁₋₅, C₂₋₆, C₃₋₆, C₄₋₆, C₅₋₆, C₂₋₅, etc.).

In general, under standard nomenclature rules used throughout thisdisclosure, the terminal portion of the designated side chain isdescribed first followed by the adjacent functionality toward the pointof attachment. Thus, for example, a “phenylC₁-C₆ alkylamidoC₁-C₆alkyl”substituent refers to a group of the formula:

Unless otherwise noted, it is intended that the definition of anysubstituent or variable at a particular location in a molecule beindependent of its definitions elsewhere in that molecule. It isunderstood that substituents and substitution patterns on the compoundsof this invention can be selected by one of ordinary skill in the art toprovide compounds that are chemically stable and that can be readilysynthesized by techniques known in the art as well as those methods setforth herein.

As used herein, the term “subject” as used herein, refers to an animal,preferably a mammal, most preferably a human, who has been the object oftreatment, observation or experiment.

As used herein, the term “therapeutically effective amount” as usedherein, means that amount of active compound or pharmaceutical agentthat elicits the biological or medicinal response in a tissue system,animal or human that is being sought by a researcher, veterinarian,medical doctor or other clinician, which includes alleviation or partialalleviation of the symptoms of the disease or disorder being treated.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the therapeuticallyeffective amounts, as well as any product which results, directly orindirectly, from combinations of the specified ingredients in thespecified amounts.

For the purposes of the present invention, the term “antagonist” is usedto refer to a compound capable of producing, depending on thecircumstance, a functional antagonism of the TRPM8 ion channel,including competitive antagonists, non-competitive antagonists,desensitizing agonists, and partial agonists.

For the purposes of the present invention, the term “inflammatoryhypersensitivity” is used to refer to a condition that is characterizedby one or more hallmarks of inflammation, including edema, erythema,hyperthermia and pain, and/or by an exaggerated physiologic orpathophysiologic response to one or more than one type of stimulation,including thermal, mechanical and/or chemical stimulation.

For purposes of the present invention, the term “TRPM8-modulated” isused to refer to the condition of being affected by the modulation ofthe TRPM8 receptor, including the state of being mediated by the TRPM8receptor.

An embodiment of the invention is a method of treating or preventing atleast one of the following diseases, syndromes, and conditions selectedfrom migraine, post herpetic neuralgia, post traumatic neuralgia, postchemotherapy neuralgia, complex regional pain syndrome I and II (CRPSI/II), fibromyalgia, inflammatory bowel disease, pruritis, asthma,chronic obstructive pulmonary disease, toothache, bone pain or pyresisin a mammal, which method comprises, consists of and/or consistsessentially of administering to subject, including an animal, a mammal,and a human in need of such treatment or prevention a therapeuticallyeffective amount of a TRPM8 antagonist that is a compound of Formula(I).

Another embodiment of the invention is a method of treating orpreventing at least one of the following diseases, syndromes, andconditions selected from hypertension, peripheral vascular disease,Raynaud's disease, reperfusion injury or frostbite in a mammal, whichmethod comprises administering to a subject, including an animal, amammal, and a human in need of such treatment or prevention atherapeutically effective amount of a TRPM8 antagonist that is acompound of Formula (I).

A further embodiment of the invention is a method of acceleratingpost-anesthetic recovery or post hypothermia recovery in a subject,including an animal, a mammal, and a human, which method comprisesadministering to a subject, including an animal, a mammal, and a humanin need of such accelerated recovery a therapeutically effective amountof a TRPM8 antagonist that is a compound of Formula (I).

An embodiment of the present invention is directed to compounds ofFormula (I)

wherein

-   -   a) A is phenyl, cyclopentyl, or cyclohexyl;        -   provided that when A is phenyl, R₂ is other than 2-chloro,            2-C₁₋₄alkoxy, 4-C₁₋₄alkoxy, 2-C₁₋₄alkoxycarbonyl, or            4-C₁₋₄alkoxycarbonyl;        -   or, when A is phenyl, L is —N(R₃)(CH₂)_(n)—, and R₁ and L            are attached to adjacent carbon atoms of phenyl, R₁ and R₃            are optionally taken with the atoms to which they are            attached to form dihydro-indol-1-yl or            dihydro-quinolin-1-yl;    -   b) A is phenyl;        -   provided that when A is phenyl, R₂ is other than 2-chloro,            2-C₁₋₄alkoxy, 4-C₁₋₄alkoxy, 2-C₁₋₄alkoxycarbonyl, or            4-C₁₋₄alkoxycarbonyl;        -   and, when L is —N(R₃)(CH₂)_(n)—, and R₁ and L are attached            to adjacent carbon atoms of phenyl, R₁ and R₃ are optionally            taken with the atoms to which they are attached to form            dihydro-indol-1-yl;    -   c) A is phenyl; provided that when A is phenyl, R₂ is other than        2-chloro, 2-C₁₋₄alkoxy, 4-C₁₋₄alkoxy, 2-C₁₋₄alkoxycarbonyl, or        4-C₁₋₄alkoxycarbonyl;    -   d) R₁ is one to two substituents selected from the group        consisting of hydrogen, C₁₋₄alkyl, C₁₋₄alkoxy, fluoro, chloro,        and hydroxy;    -   e) R₁ is one to two substituents selected from the group        consisting of hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, fluoro, and        chloro;    -   f) R₁ is one to two substituents selected from the group        consisting of hydrogen, C₁₋₃alkyl, methoxy, fluoro, and chloro;    -   g) R₁ is one to two substituents selected from the group        consisting of hydrogen, methyl, fluoro, and chloro;    -   h) L is —Z—(CH₂)_(n)— or —CH₂—; wherein n is 0 or 1; and Z is S        or NR₃; provided that when Z is S, A is phenyl;    -   i) L is —Z—(CH₂)_(n)—; wherein n is 0; and Z is S or NR₃;        provided that when Z is S, A is phenyl;    -   j) L is —Z—(CH₂)_(n)—; wherein n is 0; and Z is NR₃;    -   k) R₂ is hydrogen, C₁₋₄alkyl, trifluoromethyl, chloro, or bromo;        provided that when A is phenyl, R₂ is other than 2-chloro;    -   l) R₂ is t-butyl, trifluoromethyl, chloro, or bromo; provided        that when A is phenyl, R₂ is other than 2-chloro;    -   m) A is phenyl and R₂ is 4-t-butyl, 4-trifluoromethyl, or        4-chloro;    -   n) R₃ is hydrogen or methyl;    -   o) R₃ is hydrogen;    -   p) R₄ is hydrogen;    -   q) R₅ is indanyl or a 9 to 10-membered heteroaryl selected from        the group consisting of benzimidazolyl, indolyl, quinolinyl,        isoquinolinyl, and 1H-indazolyl; wherein R₅ is optionally        substituted with one to two substituents independently selected        from the group consisting of C₁₋₄alkyl, C₁₋₄alkoxy, and fluoro;    -   r) R₅ is a 9 to 10-membered heteroaryl selected from the group        consisting of indolyl, quinolinyl, isoquinolinyl, and        1H-indazolyl; wherein R₅ is optionally substituted with 1        C₁₋₄alkyl substituent;    -   s) R₅ is a 9 to 10-membered heteroaryl selected from the group        consisting of quinolin-5-yl, isoquinolin-5-yl, and 1H-indazolyl;    -   and enantiomers, diastereomers, solvates and pharmaceutically        acceptable salts thereof;

-   provided that a compound of Formula (I) is other than

-   a compound wherein A is phenyl, R₁ is 3-chloro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, n is 1, Z is NR₃, R₃ is    hydrogen, R₄ is hydrogen, and R₅ is isoquinolin-5-yl;

-   a compound wherein A is phenyl, R₁ is 2-fluoro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ is    hydrogen, R₄ is hydrogen, and R₅ is indol-4-yl;

-   and

-   a compound wherein A is phenyl, R₁ is 4-methoxy, R₂ is 4-chloro, L    is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ is hydrogen, R₄ is hydrogen,    and R₅ is isoquinolin-5-yl.

-   and any combination of embodiments a) through s) above, provided    that it is understood that combinations in which different    embodiments of the same substituent would be combined are excluded.

A further embodiment of the present invention is directed to a compoundof Formula (I)

wherein

-   A is phenyl, cyclopentyl, or cyclohexyl;-   provided that when A is phenyl, R₂ is other than 2-chloro;-   or, when A is phenyl, L is —N(R₃)(CH₂)_(n)—, and R₁ and L are    attached to adjacent carbon atoms, R₁ and R₃ are optionally taken    with the atoms to which they are attached to form dihydro-indol-1-yl    or dihydro-quinolin-1-yl;-   R₁ is one to two substituents selected from the group consisting of    hydrogen, C₁₋₄alkyl, C₁₋₄alkoxy, fluoro, chloro, and hydroxy;-   L is —Z—(CH₂)_(n)— or —CH₂—; wherein n is 0 or 1; and Z is S or NR₃;    provided that when Z is S, A is phenyl-   R₂ is hydrogen, C₁₋₄alkyl, trifluoromethyl, chloro, or bromo;-   R₃ is hydrogen or methyl;-   R₄ is hydrogen;-   and-   R₅ is indanyl or a 9 to 10-membered heteroaryl selected from the    group consisting of benzimidazolyl, indolyl, quinolinyl,    isoquinolinyl, and 1H-indazolyl; wherein R₅ is optionally    substituted with 1-2 substituents independently selected from the    group consisting of C₁₋₄alkyl, C₁₋₄alkoxy, and fluoro;-   and enantiomers, diastereomers, solvates and pharmaceutically    acceptable salts thereof;-   provided that a compound of Formula (I) is other than-   a compound wherein A is phenyl, R₁ is 3-chloro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, n is 1, Z is NR₃, R₃ is    hydrogen, R₄ is hydrogen, and R₅ is isoquinolin-5-yl;-   a compound wherein A is phenyl, R₁ is 2-fluoro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ is    hydrogen, R₄ is hydrogen, and R₅ is indol-4-yl;-   and-   a compound wherein A is phenyl, R₁ is 4-methoxy, R₂ is 4-chloro, L    is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ is hydrogen, R₄ is hydrogen,    and R₅ is isoquinolin-5-yl.

A further embodiment of the present invention is directed to a compoundof Formula (I)

-   Formula (I) wherein-   A is phenyl;-   provided that when A is phenyl, R₂ is other than 2-chloro;-   or, when A is phenyl, L is —N(R₃)(CH₂)_(n)—, and R₁ and L are    attached to adjacent carbon atoms, R₁ and R₃ are optionally taken    with the atoms to which they are attached to form    dihydro-indol-1-yl;-   R₁ is one to two substituents selected from the group consisting of    hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, fluoro, and chloro;-   L is —Z—(CH₂)_(n)—; wherein n is 0; and Z is S or NR₃;-   R₂ is t-butyl, trifluoromethyl, chloro, or bromo;-   R₃ is hydrogen;-   R₄ is hydrogen;-   and-   R₅ is a 9 to 10-membered heteroaryl selected from the group    consisting of indolyl, quinolinyl, isoquinolinyl, and 1H-indazolyl;    wherein R₅ is optionally substituted with 1-2 C₁₋₄alkyl    substituents;-   and enantiomers, diastereomers, solvates and pharmaceutically    acceptable salts thereof;-   provided that a compound of Formula (I) is other than-   a compound wherein A is phenyl, R₁ is 2-fluoro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ is    hydrogen, R₄ is hydrogen, and R₅ is indol-4-yl;-   and-   a compound wherein A is phenyl, R₁ is 4-methoxy, R₂ is 4-chloro, L    is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ is hydrogen, R₄ is hydrogen,    and R₅ is isoquinolin-5-yl.

A further embodiment of the present invention is directed to a compoundof Formula (I)

wherein

-   A is phenyl; provided that R₂ is other than 2-chloro;-   R₁ is one to two substituents selected from the group consisting of    hydrogen, C₁₋₃alkyl, methoxy, fluoro, and chloro;-   L is —Z—(CH₂)_(n)—; wherein n is 0; and Z is NR₃;-   R₂ is t-butyl, trifluoromethyl, chloro, or bromo;-   R₃ and R₄ are each hydrogen;-   and-   R₅ is a 9 to 10-membered heteroaryl selected from the group    consisting of quinolinyl, isoquinolinyl, and 1H-indazolyl;-   and enantiomers, diastereomers, solvates and pharmaceutically    acceptable salts thereof;-   provided that a compound of Formula (I) is other than-   a compound wherein A is phenyl, R₁ is 4-methoxy, R₂ is 4-chloro, L    is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ is hydrogen, R₄ is hydrogen,    and R₅ is isoquinolin-5-yl.-   and enantiomers, diastereomers, racemates, and pharmaceutically    acceptable salts thereof.

A further embodiment of the present invention is directed to a compoundof Formula (I)

wherein

-   A is phenyl;-   R₁ is one to two substituents selected from the group consisting of    hydrogen, methyl, fluoro, and chloro;-   L is —Z—(CH₂)_(n)—; wherein n is 0; and Z is NR₃;-   R₂ is 4-t-butyl, 4-trifluoromethyl, or 4-chloro;-   R₃ and R₄ are each hydrogen;-   and-   R₅ is a 9 to 10-membered heteroaryl selected from the group    consisting of quinolin-5-yl, isoquinolin-5-yl, and 1H-indazolyl;-   and enantiomers, diastereomers, racemates, and pharmaceutically    acceptable salts thereof.

A further embodiment of the present invention is directed to compoundsof Formula (Ia) wherein L is —Z—(CH₂)_(n)—, and R₄ is hydrogen;stereocenter 1 is defined hereinbelow, and, in certain instances when R₅contains a stereocenter, the stereocenter is defined as stereocenter 2.

selected from the group consisting of

TABLE 1 Cpd Center No. A R₁ R₂ n Z R₃ R₅ (1, 2) 1 phenyl H 4-t-butyl 0NR₃ H isoquinolin-5-yl 2 phenyl t-butyl H 0 NR₃ H isoquinolin-5-yl 3phenyl H 4-t-butyl 0 NR₃ methyl isoquinolin-5-yl 4 phenyl H 4- 0 NR₃ Hisoquinolin-5-yl trifluoro methyl 5 phenyl H 4-t-butyl 0 NR₃ H1H-indazol-4-yl 6 phenyl H 4-t-butyl 0 NR₃ H quinolin-6-yl 7 phenyl H4-t-butyl 0 NR₃ H indol-4-yl 8 phenyl H 4-t-butyl 0 NR₃ Hisoquinolin-6-yl 9 phenyl H 4-t-butyl 0 NR₃ H 1-methyl-1H-benzoimidazol- 4-yl 10 phenyl H 4- 0 NR₃ H indan-1-yl (RS, S) trifluoromethyl 11 phenyl 2-chloro 4- 0 NR₃ H isoquinolin-5-yl trifluoro methyl12 phenyl 3-chloro 4- 0 NR₃ H isoquinolin-5-yl trifluoro methyl 13phenyl 4-chloro 4- 0 NR₃ H isoquinolin-5-yl trifluoro methyl 14 phenyl2-chloro 4- 1 NR₃ H isoquinolin-5-yl trifluoro methyl 15 phenyl 4-chloro4- 1 NR₃ H isoquinolin-5-yl trifluoro methyl 16 cyclo H 4- 0 NR₃ Hisoquinolin-5-yl pentyl trifluoro methyl 17 cyclohexyl H 4- 0 NR₃ Hisoquinolin-5-yl trifluoro methyl 18 phenyl taken with 4- 0 NR₃ —isoquinolin-5-yl R₃ to form trifluoro 2,3- methyl dihydro- indol-1-yl 19phenyl taken with 4- 0 NR₃ — isoquinolin-5-yl R₃ to form trifluoro 3,4-methyl dihydro- 2H- quinolin-1- yl 20 phenyl H 4-t-butyl 0 NR₃ H1H-indazol-7-yl 21 phenyl 2-methoxy 4- 0 NR₃ H isoquinolin-5-yltrifluoro methyl 22 phenyl 2-methyl 4- 0 NR₃ H isoquinolin-5-yltrifluoro methyl 23 phenyl 2- 4- 0 NR₃ H isoquinolin-5-yl isopropyltrifluoro methyl 24 phenyl 2-fluoro 4- 0 NR₃ H isoquinolin-5-yltrifluoro methyl 25 phenyl 2,6- 4- 0 NR₃ H isoquinolin-5-yl dimethyltrifluoro methyl 26 phenyl taken with 4- 0 NR₃ — isoquinolin-5-yl R₃ toform trifluoro 7-methyl- methyl 2,3- dihydro- indol-1-yl 27 phenyl H 4-0 NR₃ H 1H-indazol-4-yl trifluoro methyl 28 phenyl H 4- 0 NR₃ H1H-indazol-7-yl trifluoro methyl 29 phenyl H 4- 0 NR₃ H quinolin-8-yltrifluoro methyl 30 phenyl H 4- 0 NR₃ H isoquinolin-5-yl 1 * S trifluoromethyl 31 phenyl H 4- 0 NR₃ H isoquinolin-5-yl 1 * R trifluoro methyl 32phenyl H 4-bromo 0 NR₃ H isoquinolin-5-yl 33 phenyl H 4-bromo 0 NR₃ Hindol-4-yl 34 phenyl 4-fluoro 4- 0 NR₃ H indol-4-yl trifluoro methyl 35phenyl 4-fluoro 4- 0 NR₃ H isoquinolin-5-yl trifluoro methyl 36 phenyl3-fluoro 4- 0 NR₃ H isoquinolin-5-yl trifluoro methyl 37 phenyl 3-fluoro4- 0 NR₃ H indol-4-yl trifluoro methyl 38 phenyl H 4-chloro 0 NR₃ Hindol-4-yl 39 phenyl H 4-chloro 0 NR₃ H isoquinolin-5-yl 40 phenyl2-fluoro 4-bromo 0 NR₃ H indol-4-yl 41 phenyl 2-fluoro 4-bromo 0 NR₃ Hisoquinolin-5-yl 42 phenyl H 3- 0 NR₃ H isoquinolin-5-yl trifluoromethyl 43 phenyl H 3- 0 NR₃ H indol-4-yl trifluoro methyl 44 phenyl2-fluoro 4-chloro 0 NR₃ H isoquinolin-5-yl 45 phenyl 4-fluoro 4-chloro 0NR₃ H isoquinolin-5-yl 46 phenyl 2,4- 4-chloro 0 NR₃ H isoquinolin-5-yldifluoro 47 phenyl 4-methoxy 4-chloro 0 NR₃ H quinolin-5-yl 48 phenyl2,6- 4-chloro 0 NR₃ H isoquinolin-5-yl difluoro 49 phenyl 2,6- 4-chloro0 NR₃ H quinolin-5-yl difluoro 50 phenyl 2,4- 4- 0 NR₃ Hisoquinolin-5-yl difluoro trifluoro methyl 51 phenyl 2,3- 4-chloro 0 NR₃H isoquinolin-5-yl difluoro 52 phenyl 2,3- 4-chloro 0 NR₃ Hquinolin-5-yl difluoro 53 phenyl 2,5- 4-chloro 0 NR₃ H isoquinolin-5-yldifluoro 54 phenyl 2,5- 4-chloro 0 NR₃ H quinolin-5-yl difluoro 56phenyl H 4- 0 S H isoquinolin-5-yl trifluoro methyl

A further embodiment of the present invention is directed to compoundsof Formula (Ib) wherein L is —CH₂—, and R₄ is hydrogen

selected from the group consisting of

TABLE 2 Cpd No. A R₁ R₂ n R₅ 55 phenyl 2-fluoro 4- 0 isoquinolin-5-yltrifluoro methyl

A further embodiment of the present invention is directed to a compoundof Formula (I)

selected from the group consisting of

-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-t-butyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is t-butyl, R₂ is    H, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and R₅ is    isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-t-butyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is methyl, n is 0, R₄ is    H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-t-butyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is 1H-indazol-4-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-t-butyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is quinolin-6-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-t-butyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is indol-4-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-t-butyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is isoquinolin-6-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-t-butyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is 1-methyl-1H-benzoimidazol-4-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is indan-1-yl (RS, S);-   a compound of Formula (I) wherein A is phenyl, R₁ is 2-chloro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 3-chloro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 4-chloro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2-chloro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 1, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 4-chloro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 1, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is cyclopentyl, R₁ is H, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is cyclohexyl, R₁ is H, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is taken with R₃    to form 2,3-dihydro-indol-1-yl, R₂ is 4-trifluoromethyl, L is    —Z—(CH₂)_(n)—, Z is NR₃, n is 0, R₄ is H, and R₅ is    isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is taken with R₃    to form 3,4-dihydro-2H-quinolin-1-yl, R₂ is 4-trifluoromethyl, L is    —Z—(CH₂)_(n)—, Z is NR₃, n is 0, R₄ is H, and R₅ is    isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-t-butyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is 1H-indazol-7-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2-methoxy, R₂    is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0,    R₄ is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2-methyl, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2-isopropyl, R₂    is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0,    R₄ is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2-fluoro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2,6-dimethyl,    R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is    0, R₄ is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is taken with R₃    to form 7-methyl-2,3-dihydro-indol-1-yl, R₂ is 4-trifluoromethyl, L    is -Z-(CH₂)_(n)—, Z is NR₃, n is 0, R₄ is H, and R₅ is    isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is 1H-indazol-4-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is 1H-indazol-7-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is quinolin-8-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl (*S);-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl (*R);-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-bromo, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and    R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-bromo, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and    R₅ is indol-4-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 4-fluoro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is indol-4-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 4-fluoro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 3-fluoro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 3-fluoro, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is indol-4-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is indol-4-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2-fluoro, R₂ is    4-bromo, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and    R₅ is indol-4-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2-fluoro, R₂ is    4-bromo, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and    R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    3-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    3-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄    is H, and R₅ is indol-4-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2-fluoro, R₂ is    4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 4-fluoro, R₂ is    4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2,4-difluoro,    R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is    H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 4-methoxy, R₂    is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,    and R₅ is quinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2,6-difluoro,    R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is    H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2,6-difluoro,    R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is    H, and R₅ is quinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2,4-difluoro,    R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is    0, R₄ is H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2,3-difluoro,    R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is    H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2,3-difluoro,    R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is    H, and R₅ is quinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2,5-difluoro,    R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is    H, and R₅ is isoquinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2,5-difluoro,    R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is    H, and R₅ is quinolin-5-yl;-   a compound of Formula (I) wherein A is phenyl, R₁ is 2-fluoro, R₂ is    4-trifluoromethyl, L is —CH₂—, R₄ is H, and R₅ is isoquinolin-5-yl;-   and-   a compound of Formula (I) wherein A is phenyl, R₁ is H, R₂ is    4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is S, R₃ is H, n is 0, R₄    is H, and R₅ is isoquinolin-5-yl;-   and enantiomers, diastereomers, solvates and pharmaceutically    acceptable salts thereof.

For use in medicine, salts of compounds of formula (I) refer tonon-toxic “pharmaceutically acceptable salts.” Other salts may, however,be useful in the preparation of compounds of formula (I) or of theirpharmaceutically acceptable salts thereof. Suitable pharmaceuticallyacceptable salts of compounds of formula (I) include acid addition saltswhich can, for example, be formed by mixing a solution of the compoundwith a solution of a pharmaceutically acceptable acid, such as,hydrochloric acid, sulfuric acid, fumaric acid, maleic acid, succinicacid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonicacid or phosphoric acid.

Furthermore, where the compounds of formula (I) carry an acidic moiety,suitable pharmaceutically acceptable salts thereof may include alkalimetal salts, such as, sodium or potassium salts; alkaline earth metalsalts, such as, calcium or magnesium salts; and salts formed withsuitable organic ligands, such as, quaternary ammonium salts. Thus,representative pharmaceutically acceptable salts include acetate,benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate,bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate,citrate, dihydrochloride, edetate, edisylate, estolate, esylate,fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate,hexylresorcinate, hydrabamine, hydrobromide, hydrochloride,hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate,malate, maleate, mandelate, mesylate, methylbromide, methylnitrate,methylsulfate, mucate, napsylate, nitrate, N-methylglucamine ammoniumsalt, oleate, pamoate (embonate), palmitate, pantothenate,phosphate/diphosphate, polygalacturonate, salicylate, stearate, sulfate,subacetate, succinate, tannate, tartrate, teoclate, tosylate,triethiodide and valerate.

Representative acids and bases that may be used in the preparation ofpharmaceutically acceptable salts include acids including acetic acid,2,2-dichloroactic acid, acylated amino acids, adipic acid, alginic acid,ascorbic acid, L-aspartic acid, benzenesulfonic acid, benzoic acid,4-acetamidobenzoic acid, (+)-camphoric acid, camphorsulfonic acid,(+)-(1S)-camphor-10-sulfonic acid, capric acid, caproic acid, caprylicacid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid,ethane-1,2-disulfonic acid, ethanesulfonic acid,2-hydroxy-ethanesulfonic acid, formic acid, fumaric acid, galactaricacid, gentisic acid, glucoheptonic acid, D-gluconic acid, D-glucoronicacid, L-glutamic acid, α-oxo-glutaric acid, glycolic acid, hippuricacid, hydrobromic acid, hydrochloric acid, (+)-L-lactic acid,(±)-DL-lactic acid, lactobionic acid, maleic acid, (−)-L-malic acid,malonic acid, (±)-DL-mandelic acid, methanesulfonic acid,naphthalene-2-sulfonic acid, naphthalene-1,5-disulfonic acid,1-hydroxy-2-naphthoic acid, nicotinic acid, nitric acid, oleic acid,orotic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid,L-pyroglutamic acid, salicylic acid, 4-amino-salicylic acid, sebaicacid, stearic acid, succinic acid, sulfuric acid, tannic acid,(+)-L-tartaric acid, thiocyanic acid, p-toluenesulfonic acid andundecylenic acid;

and bases including ammonia, L-arginine, benethamine, benzathine,calcium hydroxide, choline, deanol, diethanolamine, diethylamine,2-(diethylamino)-ethanol, ethanolamine, ethylenediamine,N-methyl-glucamine, hydrabamine, 1H-imidazole, L-lysine, magnesiumhydroxide, 4-(2-hydroxyethyl)-morpholin, piperazine, potassiumhydroxide, 1-(2-hydroxyethyl)-pyrrolidine, secondary amine, sodiumhydroxide, triethanolamine, tromethamine and zinc hydroxide.

Embodiments of the present invention include prodrugs of compounds offormula (I). In general, such prodrugs will be functional derivatives ofthe compounds that are readily convertible in vivo into the requiredcompound. Thus, in the methods of treating or preventing embodiments ofthe present invention, the term “administering” encompasses thetreatment or prevention of the various diseases, conditions, syndromesand disorders described with the compound specifically disclosed or witha compound that may not be specifically disclosed, but which converts tothe specified compound in vivo after administration to a patient.Conventional procedures for the selection and preparation of suitableprodrug derivatives are described, for example, in “Design of Prodrugs”,ed. H. Bundgaard, Elsevier, 1985.

Where the compounds according to embodiments of this invention have atleast one chiral center, they may accordingly exist as enantiomers.Where the compounds possess two or more chiral centers, they mayadditionally exist as diastereomers. It is to be understood that allsuch isomers and mixtures thereof are encompassed within the scope ofthe present invention. Furthermore, some of the crystalline forms forthe compounds may exist as polymorphs and as such are intended to beincluded in the present invention. In addition, some of the compoundsmay form solvates with water (i.e., hydrates) or common organicsolvents, and such solvates are also intended to be encompassed withinthe scope of this invention. The skilled artisan will understand thatthe term compound as used herein, is meant to include solvated compoundsof Formula I.

Where the processes for the preparation of the compounds according tocertain embodiments of the invention give rise to a mixture ofstereoisomers, these isomers may be separated by conventionaltechniques, such as, preparative chromatography. The compounds may beprepared in racemic form, or individual enantiomers may be preparedeither by enantiospecific synthesis or by resolution. The compounds may,for example, be resolved into their component enantiomers by standardtechniques, such as, the formation of diastereomeric pairs by saltformation with an optically active acid, such as,(−)-di-p-toluoyl-d-tartaric acid and/or (+)-di-p-toluoyl-l-tartaric acidfollowed by fractional crystallization and regeneration of the freebase. The compounds may also be resolved by formation of diastereomericesters or amides, followed by chromatographic separation, and removal ofthe chiral auxiliary. Alternatively, the compounds may be resolved usinga chiral HPLC column.

One embodiment of the present invention is directed to a composition,including a pharmaceutical composition, comprising, consisting of,and/or consisting essentially of the (+)-enantiomer of a compound offormula (I) wherein said composition is substantially free from the(−)-isomer of said compound. In the present context, substantially freemeans less than about 25%, preferably less than about 10%, morepreferably less than about 5%, even more preferably less than about 2%and even more preferably less than about 1% of the (−)-isomer calculatedas.

${\%\mspace{14mu}( + )\text{-}{enantiomer}} = {\frac{\left( {{{mass}( + )}\text{-}{enantiomer}} \right)}{\begin{matrix}{\left( {{mass}( + )\text{-}{enantiomer}} \right) +} \\\left( {{{mass}( - )}\text{-}{enantiomer}} \right)\end{matrix}} \times 100}$

Another embodiment of the present invention is a composition, includinga pharmaceutical composition, comprising, consisting of, and consistingessentially of the (−)-enantiomer of a compound of formula (I) whereinsaid composition is substantially free from the (+)-isomer of saidcompound. In the present context, substantially free from means lessthan about 25%, preferably less than about 10%, more preferably lessthan about 5%, even more preferably less than about 2% and even morepreferably less than about 1% of the (+)-isomer calculated as

${\%\mspace{14mu}( - )\text{-}{enantiomer}} = {\frac{\left( {{{mass}( - )}\text{-}{enantiomer}} \right)}{\begin{matrix}{\left( {{mass}( + )\text{-}{enantiomer}} \right) +} \\\left( {{{mass}( - )}\text{-}{enantiomer}} \right)\end{matrix}} \times 100.}$

During any of the processes for preparation of the compounds of thevarious embodiments of the present invention, it may be necessary and/ordesirable to protect sensitive or reactive groups on any of themolecules concerned. This may be achieved by means of conventionalprotecting groups, such as those described in Protective Groups inOrganic Chemistry, ed. J. F. W. McOmie, Plenum Press, 1973; and T. W.Greene & P. G. M. Wuts, Protective Groups in Organic Synthesis, JohnWiley & Sons, 1991. The protecting groups may be removed at a convenientsubsequent stage using methods known from the art.

Even though the compounds of embodiments of the present invention(including their pharmaceutically acceptable salts and pharmaceuticallyacceptable solvates) can be administered alone, they will generally beadministered in admixture with a pharmaceutically acceptable carrier, apharmaceutically acceptable excipient and/or or a pharmaceuticallyacceptable diluent selected with regard to the intended route ofadministration and standard pharmaceutical practice. Thus, particularembodiments of the present invention are directed to pharmaceuticalcompositions comprising consisting of, and consisting essentially ofcompounds of formula (I) and at least one pharmaceutically acceptablecarrier, pharmaceutically acceptable excipient, and/or pharmaceuticallyacceptable diluent.

By way of example, in the pharmaceutical compositions of embodiments ofthe present invention, the compounds of formula (I) may be admixed withany suitable binder(s), lubricant(s), suspending agent(s), coatingagent(s), r solubilizing agent(s).

Solid oral dosage forms, such tablets or capsules, containing thecompounds of the present invention may be administered at least onedosage form at a time, as appropriate. It is also possible to administerthe compounds in sustained release formulations.

Additional oral forms in which the present inventive compounds may beadministered include elixirs, solutions, syrups, and suspensions; eachoptionally containing flavoring agents and coloring agents.

Alternatively, compounds of formula (I) can be administered byinhalation (intratracheal or intranasal) or in the form of a suppositoryor pessary, or they may be applied topically in the form of a lotion,solution, cream, ointment or dusting powder. For example, they can beincorporated into a cream comprising, consisting of, and/or consistingessentially of an aqueous emulsion of polyethylene glycols or liquidparaffin. They can also be incorporated, at a concentration of betweenabout 1% and about 10% by weight of the cream, into an ointmentcomprising, consisting of, and/or consisting essentially of a white waxor white soft paraffin base together with any stabilizers andpreservatives as may be required. An alternative means of administrationinclude transdermal administration by using a skin or transdermal patch

The pharmaceutical compositions of the present invention (as well as thecompounds of the present invention alone) can also be injectedparenterally, for example, intracavernosally, intravenously,intramuscularly, subcutaneously, intradermally or intrathecally. In thiscase, the compositions will also include at least one of a suitablecarrier, a suitable excipient, and a suitable diluent.

For parenteral administration, the pharmaceutical compositions of thepresent invention are best used in the form of a sterile aqueoussolution that may contain other substances, for example, enough salts ormonosaccharides to make the solution isotonic with blood.

For buccal or sublingual administration, the pharmaceutical compositionsof the present invention may be administered in the form of tablets orlozenges, which can be formulated in a conventional manner.

By way of further example, pharmaceutical compositions containing atleast one of the compounds of formula (I) as the active ingredient canbe prepared by mixing the compound(s) with a pharmaceutically acceptablecarrier, a pharmaceutically acceptable diluent, and/or apharmaceutically acceptable excipient according to conventionalpharmaceutical compounding techniques.

The carrier, excipient, and diluent may take a wide variety of formsdepending upon the desired route of administration (e.g., oral,parenteral, etc.). Thus for liquid oral preparations, such as,suspensions, syrups, elixirs and solutions, suitable carriers,excipients, and diluents include water, glycols, oils, alcohols,flavoring agents, preservatives, stabilizers, coloring agents and thelike; for solid oral preparations, such as, powders, capsules andtablets, suitable carriers, excipients, and diluents include starches,sugars, diluents, granulating agents, lubricants, binders,disintegrating agents and the like. Solid oral preparations also may beoptionally coated with substances, such as, sugars, or beenterically-coated so as to modulate the major site of absorption anddisintegration. For parenteral administration, the carrier, excipient,and/or diluent will usually include sterile water, and other ingredientsmay be added to increase solubility or preservation of the composition.Injectable suspensions or solutions may also be prepared utilizingaqueous carriers along with appropriate carriers, excipients, and/ordiluents, such as solubilizers and preservatives.

A therapeutically effective amount of a compound of formula (I) or apharmaceutical composition thereof includes a dose range of from about0.1 mg to about 3000 mg, in particular from about 1 mg to about 1000 mgor, more particularly, from about 10 mg to about 500 mg of activeingredient in a regimen of about 1 to 4 times per day for an average (70kg) human; although, it is apparent to one skilled in the art that thetherapeutically effective amount for active compounds of the inventionwill vary as will the conditions being treated.

For oral administration, a pharmaceutical composition is preferablyprovided in the form of tablets containing about 0.01, about 10.0, about50.0, about 100, about 150, about 200, about 250, and about 500milligrams of the active ingredient.

Advantageously, a compound of formula (I) may be administered in asingle daily dose, or the total daily dosage may be administered individed doses of two, three or four times daily.

Optimal dosages of a compound of formula (I) to be administered may bereadily determined and will vary with the particular compound used, themode of administration, the strength of the preparation, and theadvancement of the disease, syndrome, condition, or disorder. Inaddition, factors associated with the particular subject being treated,including subject age, weight, diet and time of administration, willresult in the need to adjust the dose to achieve an appropriatetherapeutic level. The above dosages are thus exemplary of the averagecase. There can be, of course, individual instances wherein higher orlower dosage ranges are merited, and such are within the scope of thisinvention.

Compounds of formula (I) may be administered in any of the foregoingcompositions and dosage regimens or by means of those compositions anddosage regimens established in the art whenever use of a compound offormula (I) is required for a subject in need thereof.

As antagonists of the TRPM8 ion channel, the compounds of formula (I)are useful in methods for treating or preventing a disease, syndrome orcondition in a subject, including an animal, a mammal and a human inwhich the disease, syndrome, or condition is affected by the modulationof TRPM8 receptors. Such methods comprise, consist of, and/or consistessentially of administering to a subject, including an animal, a mammaland a human in need of such treatment or prevention a therapeuticallyeffective amount of a compound, salt or solvate of formula (I). Inparticular, the compounds of formula (I) are useful for preventing ortreating—pain, or diseases, syndromes, or conditions, causing such pain,or pulmonary or vascular dysfunction. More particularly, the compoundsof formula (I) are useful for preventing or treating inflammatory pain,inflammatory hypersensitivity conditions, neuropathic pain, anxiety,depression, and cardiovascular disease aggravated by cold, includingperipheral vascular disease, vascular hypertension, pulmonaryhypertension, Raynaud's disease, and coronary artery disease, byadministering to a subject in need thereof a therapeutically effectiveamount of a compound of formula (I).

Examples of inflammatory pain include pain due to a disease, conditionsyndrome, disorder or pain state, including inflammatory bowel disease,visceral pain, migraine, post operative pain, osteoarthritis, rheumatoidarthritis, back pain, lower back pain, joint pain, abdominal pain, chestpain, labor, musculoskeletal diseases, skin diseases, toothache,pyresis, burn, sunburn, snake bite, venomous snake bite, spider bite,insect sting, neurogenic bladder, interstitial cystitis, urinary tractinfection, rhinitis, contact dermatitis/hypersensitivity, itch, eczema,pharyngitis, mucositis, enteritis, irritable bowel syndrome,cholecystitis, pancreatitis, postmastectomy pain syndrome, menstrualpain, endometriosis, sinus headache, tension headache, or arachnoiditis.

One type of inflammatory pain is inflammatory hyperalgesia, which can befurther distinguished as inflammatory somatic hyperalgesia orinflammatory visceral hyperalgesia. Inflammatory somatic hyperalgesiacan be characterized by the presence of an inflammatory hyperalgesicstate in which a hypersensitivity to thermal, mechanical and/or chemicalstimuli exists. Inflammatory visceral hyperalgesia can also becharacterized by the presence of an inflammatory hyperalgesic state, inwhich an enhanced visceral irritability exists.

Examples of inflammatory hyperalgesia include a disease, syndrome,condition, disorder or pain state including inflammation,osteoarthritis, rheumatoid arthritis, back pain, joint pain, abdominalpain, musculoskeletal diseases, skin diseases, post operative pain,headaches, toothache, burn, sunburn, insect sting, neurogenic bladder,urinary incontinence, interstitial cystitis, urinary tract infection,cough, asthma, chronic obstructive pulmonary disease, rhinitis, contactdermatitis/hypersensitivity, itch, eczema, pharyngitis, enteritis,irritable bowel syndrome, and inflammatory bowel diseases includingCrohn's Disease, and or ulcerative colitis.

One embodiment of the present invention is directed to a method fortreating inflammatory somatic hyperalgesia in which a hypersensitivityto thermal, mechanical and/or chemical stimuli exists, comprising,consisting of, and/or consisting essentially of the step ofadministering to a subject in need of such treatment a therapeuticallyeffective amount of a compound, salt or solvate of formula (I).

A further embodiment of the present invention is directed to a methodfor treating neuropathic cold allodynia in which a hypersensitivity to acooling stimuli exists, comprising, consisting of, and/or consistingessentially of the step of administering to a subject in need of suchtreatment a therapeutically effective amount of a compound, salt orsolvate of formula (I).

A further embodiment of the present invention is directed to a methodfor treating neuropathic cold allodynia in which a hypersensitivity to acooling stimuli exists, comprising, consisting of, and/or consistingessentially of the step of administering to a subject in need of suchtreatment a therapeutically effective amount of a compound, salt orsolvate of formula (I).

Examples of an inflammatory hypersensitivity condition include urinaryincontinence, benign prostatic hypertrophy, cough, asthma, rhinitisand/or nasal hypersensitivity, itch, contact dermatitis and/or dermalallergy, and chronic obstructive pulmonary disease.

Examples of a neuropathic pain include pain due to a disease, syndrome,condition, disorder, or pain state, include cancer, neurologicaldisorders, spine and peripheral nerve surgery, brain tumors, traumaticbrain injury (TBI), spinal cord trauma, chronic pain syndromes,fibromyalgia, chronic fatigue syndrome, neuralgias (trigeminalneuralgia, glossopharyngeal neuralgia, postherpetic neuralgia andcausalgia), lupus, sarcoidosis, peripheral neuropathy, bilateralperipheral neuropathy, diabetic neuropathy, central pain, neuropathiesassociated with spinal cord injury, stroke, amyotrophic lateralsclerosis (ALS), Parkinson's disease, multiple sclerosis, sciaticneuritis, mandibular joint neuralgia, peripheral neuritis, polyneuritis,stump pain, phantom limb pain, bony fractures, oral neuropathic pain,Charcot's pain, complex regional pain syndrome I and II (CRPS I/II),radiculopathy, Guillain-Barre syndrome, meralgia paresthetica,burning-mouth syndrome, optic neuritis, postfebrile neuritis, migratingneuritis, segmental neuritis, Gombault's neuritis, neuronitis,cervicobrachial neuralgia, cranial neuralgia, geniculate neuralgia,glossopharyngial neuralgia, migrainous neuralgia, idiopathic neuralgia,intercostals neuralgia, mammary neuralgia, Morton's neuralgia,nasociliary neuralgia, occipital neuralgia, red neuralgia, Sluder'sneuralgia, splenopalatine neuralgia, supraorbital neuralgia, vulvodynia,or vidian neuralgia.

One type of neuropathic pain is neuropathic cold allodynia, which can becharacterized by the presence of a neuropathy-associated allodynic statein which a hypersensitivity to cooling stimuli exists. Examples ofneuropathic cold allodynia include allodynia due to a disease,condition, syndrome, disorder, or pain state including neuropathic pain(neuralgia), pain arising from spine and peripheral nerve surgery ortrauma, traumatic brain injury (TBI), trigeminal neuralgia, postherpeticneuralgia, causalgia, peripheral neuropathy, diabetic neuropathy,central pain, stroke, peripheral neuritis, polyneuritis, complexregional pain syndrome I and II (CRPS I/II) and radiculopathy.

Examples of anxiety include social anxiety, post traumatic stressdisorder, phobias, social phobia, special phobias, panic disorder,obsessive compulsive disorder, acute stress, disorder, separationanxiety disorder, and generalized anxiety disorder.

Examples of depression include major depression, bipolar disorder,seasonal affective disorder, post natal depression, manic depression,and bipolar depression.

GENERAL SYNTHETIC METHODS

Representative compounds of the present invention can be synthesized inaccordance with the general synthetic methods described below andillustrated in the schemes and examples that follow. Since the schemesare an illustration, the invention should not be construed as beinglimited by the specific chemical reactions and specific conditionsdescribed in the schemes and examples. The various starting materialsused in the schemes and examples are commercially available or may beprepared by methods well within the skill of persons versed in the art.The variables are as defined herein and within the skill of personsversed in the art.

Abbreviations used in the instant specification, particularly theschemes and examples, are as follows:

AcCl acetyl chloride AcOH glacial acetic acid aq. aqueous Bn or Bzlbenzyl conc. concentrated DBU 1,8-diazabicyclo[5.4.0]undec-7-ene DCE1,2-dichloroethane DCM dichloromethane DEAD diethyl azodicarboxylateDIAD diisopropyl azodicarboxylate DIEA diisopropyl-ethyl amine DMAP4-(dimethylamino)pyridine DMF N,N-dimethylformamide DMSOdimethylsulfoxide DPPA diphenylphosphoryl azide ESI electron-sprayionization EtOAc ethyl acetate EtOH ethanol h hour HATU2-(1H-7-azabenzotriazol-1-yl)--1,1,3,3- tetramethyl uroniumhexafluorophosphate HBTU O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluorophosphate HEK human embryonic kidney HOBt1-hydroxybenzotriazole HPLC high performance liquid chromatography LDAlithium diisopropylamide LHMDS lithium bis(trimethylsilyl)amide Memethyl MeOH methanol MHz megahertz min minutes MPLC medium pressureliquid chromatography MS mass spectroscopy NaHMDS sodiumbis(trimethylsilyl)amide NBS N-bromosuccinimide NCS N-chlorosuccinimideNMR nuclear magnetic resonance NT not tested PCC pyridiniumchlorochromate Ph phenyl Pd/C palladium on activated carbon Ph₃Ptriphenylphosphine PPA polyphosphoric acid rt room temperature TEA/Et₃Ntriethylamine TFA trifluoroacetic acid THF tetrahydrofuran TLC thinlayer chromatography TMS tetramethylsilane TMSCN trimethylsilyl cyanide

Scheme A illustrates a route for the synthesis of compounds of formula(I)-A wherein L is —Z—(CH₂)_(n)—, Z is NR₃, and R₃ is H or C₁₋₃alkyl.

A compound of formula A1 is either commercially available or may beprepared by known methods, including those described in the scientificliterature. A compound of formula A1 may be converted to itscorresponding bromide of formula A2 by the action of NBS in hydrobromicacid in carbon tetrachloride, or by another brominating agent, such as,bromine in acetic acid or by carbon tetrabromide with DBU. Treatment ofa compound of formula A2 with an appropriately substituted amine offormula A3 affords a compound of formula A4. Saponification of acompound of formula A4 by the action of an alkali metal hydroxideaffords a compound of formula A5. The carboxylic acid of formula A5 maybe coupled with an appropriately substituted amine of formula A6,R₄R₅N—H, in the presence of a peptide coupling agent, such as, HATU,DCC, HBTU, and the like, and a tertiary amine to afford a compound offormula (I)-A. Compounds of formula A6 are either commercially availableor may be prepared by known methods, including those described in thescientific literature.

An alternate route to compounds of Formula (I)-A is presented in SchemeB.

A compound of formula B1 is either commercially available or may beprepared by known methods described in the scientific literature. Acompound of formula B1 may be converted to its corresponding acidchloride by the action of a chlorinating agent, such as, thionylchloride, oxalyl chloride, and the like. Subsequent treatment of theacid chloride with a conventional brominating agent affords a compoundof formula B2. Conversion to a compound of formula (I)-A may beaccomplished by reaction of the acid chloride of formula B2 with acompound of formula A6, followed by nucleophilic displacement of thebromide by a compound of formula A3.

Scheme C illustrates a route for the synthesis of compounds of formula(I)-C wherein ring A is phenyl, L is —Z—(CH₂)_(n)—, n is 0, Z is NR₃,and R₁ and R₃ are optionally taken with the atoms to which they areattached to form dihydro-indol-1-yl or dihydro-quinolin-1-yl.

A compound of formula A3 may undergo a nucleophilic displacement with anappropriately substituted amine of formula C1 to form a compound offormula C2. Saponification of a compound of formula C2 to itscorresponding carboxylic acid, followed by coupling of the carboxylicacid with a compound of formula A7, as described herein, affords acompound of formula (I)-C.

Scheme D illustrates a route for the synthesis of compounds of formula(I)-D wherein L is —Z—(CH₂)_(n)—, Z is O.

A compound of formula D1 is either commercially available or may beprepared by known methods described in the scientific literature. Acompound of formula D1 may be converted to a compound of formula D2 viaa coupling reaction between the carboxy group of D1 and the amine offormula A6 using appropriate coupling conditions as described herein.The hydroxy function of a compound of formula D2 may be coupled with acompound of formula D3 under conventional Mitsunobu coupling conditionsto afford a compound of formula (I)-D1. Compounds of formula D3 areeither commercially available or may be prepared by known methods,including those described in the scientific literature.

Compounds of formula (I)-D2 wherein n is 1 may be prepared by thealkylation of a compound of formula D1 with a compound of formula D4 toafford a compound of formula D5, which may then be coupled with acompound of formula A6 as described herein.

Scheme E illustrates a route for the synthesis of compounds of formula(1)-E wherein ring A is phenyl, L is —Z—(CH₂)_(n)— and Z is S.

A compound of formula E1 may be converted to its corresponding acidchloride by the action of a chlorinating agent, such as, thionylchloride, oxalyl chloride, and the like. Subsequent treatment of theacid chloride with a compound of formula A6 affords a compound offormula E3. Conversion to a compound of formula (1)-E may beaccomplished by reaction of the alcohol of formula E3 with achlorinating agent, such as, thionyl chloride and the like to give acompound of formula E4, followed by nucleophilic displacement of thechloride by a compound of formula E5.

Scheme F illustrates a route for the synthesis of compounds of formula(1)-F wherein L is —CH₂—.

A compound of formula A1 may be deprotonated with an organic base, suchas, lithium diisopropylamide (LDA) and alkylated with a compound offormula F1. Compounds of formula F1 are either commercially available ormay be prepared by known methods described in the scientific literature.The ester of formula F2 may be saponified and subsequently coupled witha compound of formula A6 as described herein to yield a compound offormula (1)-F.

Compounds of Formula (I) that are chiral may be separated into theirenantiomers by chromatography on a chiral stationary phase.Alternatively, basic or acidic compounds or intermediates to compoundsof the present invention may be converted to diastereomeric salts bymixture with a chiral acid or base, respectively, and resolved intotheir enantiomers by fractional crystallization.

It is generally preferred that the respective product of each processstep be separated from other components of the reaction mixture andsubjected to purification before its use as a starting material in asubsequent step. Separation techniques typically include evaporation,extraction, precipitation and filtration. Purification techniquestypically include column chromatography (Still, W. C., et. al., J. Org.Chem. 1978, 43, 2923-2925), thin-layer chromatography, crystallizationand distillation. The structures of the final products, intermediatesand starting materials are confirmed by spectroscopic, spectrometric andanalytical methods including nuclear magnetic resonance (NMR), massspectrometry (MS) and liquid chromatography (HPLC). In the descriptionsfor the preparation of compounds of this invention, ethyl ether,tetrahydrofuran and dioxane are common examples of an ethereal solvent;benzene, toluene, hexanes and heptanes are typical hydrocarbon solventsand dichloromethane and dichloroethane are representative halogenatedhydrocarbon solvents. In those cases where the product is isolated asthe acid addition salt the free base may be obtained by techniques knownto those skilled in the art. In those cases in which the product isisolated as an acid addition salt, the salt may contain one or moreequivalents of the acid. Enantiomers of the compounds of the presentinvention may be separated using chiral HPLC.

SPECIFIC EXAMPLES

Reagents were purchased from commercial sources. Microanalyses wereperformed at Quantitative Technologies, Inc., Whitehouse, N.J. and areexpressed in percentage by weight of each element per total molecularweight. Nuclear magnetic resonance (NMR) spectra for H atoms weremeasured in the indicated solvent with (TMS) as the internal standard ona Bruker Avance or Varian (300 or 400, or 500 MHz) spectrometer. Thevalues are expressed in parts per million downfield from TMS. The massspectra (MS) were determined on an Agilent spectrometer as (ESI) m/z(M+H⁺) using an electrospray technique. Optical rotations were obtainedon a Perkin-Elmer polarimeter using the sodium D line as wavelength oflight. Unless otherwise noted, the materials used in the examples wereobtained from readily available commercial suppliers or synthesized bystandard methods known to one skilled in the art of chemical synthesis.The substituent groups, which vary between examples, are H unlessotherwise noted. Where reactions were carried out in a microwavereactor, a Biotage Initiator™ was used.

Example 1

A. Bromo-(4-chloro-phenyl)-acetic acid methyl. To a solution of Compound1a (5.0 g, 27.1 mmol) in anhydrous CCl₄ (110 mL), at room temperature,was added NBS (5.3 g, 29.8 mmol) and 48% HBr (0.2 mL). The reactionmixture was refluxed for 2 h and then the mixture was cooled, filteredand evaporated in vacuo to afford Compound 1b.

B. (4-Chloro-phenyl)-(2-fluoro-phenylamino)-acetic acid methyl ester. Toa solution of Compound 1b (2.8 g, 10.6 mmol) in acetonitrile (16 mL) wasadded 2-fluoro-phenylamine (2.95 g, 26.5 mmol) at room temperature. Thereaction mixture was refluxed for 24 h and then the solvent wasevaporated in vacuo. The resultant residue was diluted with EtOAc,washed with water and dried over Na₂SO₄. The mixture was then filteredand the filtrate was concentrated in vacuo. The resulting residue waspurified by flash column chromatography (SiO₂), eluting with ahexanes-EtOAc gradient to afford Compound 1c.

C. (4-Chloro-phenyl)-(2-fluoro-phenylamino)-acetic acid. To Compound 1c(2.79 g, 9.5 mmol) dissolved in MeOH (30 mL) was added lithium hydroxidemonohydrate (2.63 g, 62.7 mmol) and H₂0 (20 mL) at room temperature. Thereaction mixture was stirred at room temperature overnight and then thesolvent was evaporated in vacuo. The resultant residue was re-dissolvedin water and the pH was adjusted to pH 2 with 2N HCl. The resultingwhite solid was collected by filtration, washed with H₂O, hexanes, anddried to afford Compound 1d.

D.2-(4-Chloro-phenyl)-2-(2-fluoro-phenylamino)-1-[2-(2-fluoro-phenyl)-pyrrolidin-1-yl]-ethanone.To a solution of Compound 1d (50.0 mg, 0.179 mmol) in CH₂Cl₂ (2 mL) wasadded isoquinolin-5-ylamine (26.0 mg, 0.179 mmol), HATU (136 mg, 0.358mmol) and triethylamine (0.25 mL, 1.79 mmol). The reaction mixture wasstirred at room temperature overnight and then the solvent wasevaporated in vacuo. The resultant residue was purified by flash columnchromatography (SiO₂), eluting with a hexanes-EtOAc gradient to affordCompound 44. MS 406 (M+H).

Following the procedure described above for Example 1 and substitutingthe appropriate reagents, starting materials and purification methodsknown to those skilled in the art, the following compounds of thepresent invention were prepared:

MS Cpd (M + 1)⁺ 1 410 2 410 3 424 4 423 5 399 6 410 7 398 8 410 9 413 20399 27 411 28 411 29 422 32 432 33 420 34 428 35 440 36 440 37 428 38376 39 388 40 438 41 450 42 422 43 410 45 406 46 424 47 418 48 424 49424 50 458 51 424 52 424 53 424 54 424

Example 2

N-Isoquinolin-5-yl-2-phenylamino-2-(4-trifluoromethyl-phenyl)-acetamide(Cpd 30 and Cpd 31). A sample of Compound 4 was separated into itsenantiomers on an ADH 15 cm column, eluting with CH₃CN at 1 mL/min togive two products: a compound assigned to Peak A (0.40 g) and a secondcompound assigned to Peak B (0.39 g). Each sample was dissolved inacetonitrile and 2.2 equivalents of 1N ethereal H chloride were added.The solids were individually collected and dried at rt for three daysunder vacuum to afford Peak A, Compound 30, (285.5 mg), and Peak B,Compound 31 (239.0 mg).

Cpd 30: MS: C₂₄H₁₈F₃N₃O: m/z422.17 (M+1); [α]_(D)=−30.6(c=0.007, MeOH).

Cpd 31: ¹H NMR (300 MHz, DMSO-d₆) δ ppm 5.98 (s, 1 H) 6.62 (t, J=7.16Hz, 1 H) 6.85 (d, J=7.91 Hz, 2 H) 7.12 (t, J=7.91 Hz, 2 H) 7.79 (d,J=7.91 Hz, 2 H) 7.92-8.07 (m, 3 H) 8.28 (d, J=7.16 Hz, 1 H) 8.36 (d,J=8.29 Hz, 1 H) 8.54 (d, J=6.78 Hz, 1 H) 8.72 (d, J=6.40 Hz, 1 H) 9.92(s, 1 H) 11.47 (s, 1 H); MS: C₂₄H₁₈F₃N₃O: m/z422.16 (M+1); [α]_(D)=+29.6(c=0.007, MeOH).

Example 3

A. 2-Bromo-2-(4-(trifluoromethyl)phenyl)acetyl chloride. To a slurry ofCompound 3a (10.21 g, 50.0 mmol) in CCl₄ (5 mL) was added SOCl₂ (14.6mL, 200.6 mmol). The reaction was heated at 65° C. for 45 min thendiluted with CCl₄ (25 mL). N-Bromo succinimide (10.70 g, 60.1 mmol) wasadded followed by 1 drop of 48% HBr. The temperature was then increasedto 85° C. for an additional 2 h. The reaction mixture was then cooled toroom temperature and diluted with hexanes (250 mL). The solids werefiltered away and the filtrate was evaporated in vacuo. The resultingresidue was dissolved in hexanes (25 mL), filtered again, andconcentrated in vacuo to give Compound 3b in quantitive yield as apeach-colored liquid (15.29 g). ¹H NMR (CDCl₃): δ 7.73-7.58 (m, 4H),5.69 (s, 1H).

B.2-(2-Fluorophenylamino)-N-(isoquinolin-5-yl)-2-(4-(trifluoromethyl)phenyl)acetamide.A solution of Compound 3b (1.90 mL, 10.1 mmol) in DCM (50 mL) was addeddropwise over a period of 18 min to a solution of 5-amino isoquinoline(1.372 g, 9.52 mmol) in DCM (50 mL). The reaction mixture was stirred atroom temperature for an additional 30 min and then quenched withsaturated NaHCO₃ solution (50 mL). The organic portion was isolated,dried over Na₂SO₄, filtered, and diluted with DCM to a volume of 100 mL.1/19^(th) of this solution by volume (0.50 mmol) was put into a vial. Tothis was added DMF (0.5 mL) followed by 2-F aniline (0.122 mL, 1.26mmol). After stirring at ambient temperature for 2 days the reactionmixture was evaporated in vacuo and the residue was purified byreverse-phase chromatography (25-95% acetonitrile/water+0.1% TFA) togive Compound 24 as a yellow powder (0.035 g). ¹H NMR (DMSO-d₆): δ 10.71(s, 1H), 9.53 (s, 1H), 8.57 (d, 1H), 8.13 (d, 1H), 8.03 (d, 1H), 7.93(d, 2H), 7.88 (d, 1H), 7.84-7.75 (m, 3H), 7.12 (dd,1H), 6.99 (t, 1H),6.77 (t,1H), 6.68 (q,1H), 5.99 (br s, 1H), 5.73 (s,1H); MS: m/z 440.1(MH⁺).

Following the procedure described above for Example 3 and substitutingthe appropriate reagents, starting materials and purification methodsknown to those skilled in the art, the following compounds of thepresent invention were prepared:

MS Cpd (M + 1)⁺ 4 422.2 11 456.1 12 456.1 13 456.1 14 470.1 15 470.1 16414.2 17 428.2 21 452.1 22 436.1 23 464.2 25 450.1

Example 4

A. 2-Hydroxy-2-(4-(trifluoromethyl)phenyl)acetyl chloride. A suspensionof Compound 4a (2.12 g, 10.0 mmol) in thionyl chloride (10 mL, 137 mmol)was heated at reflux overnight then evaporated in vacuo to affordCompound 4b which was used without further purification (2.098 g).

B.2-Hydroxy-N-(isoquinolin-5-yl)-2-(4-(trifluoromethyl)phenyl)acetamide.To a solution of 5-aminoisoquinoline (1.153 g, 8.00 mmol) in CH₃CN (80mL) was added Compound 4b (2.09 g, 8.76 mmol) dropwise. After 4 hourspyrrolidine (2.0 mL, 24 mmol) was added. After stirring for 3 more daysthe reaction mixture was diluted with 500 mL water and extracted threetimes with 100 mL EtOAc. The combined organics were washed once with 50mL brine, dried with Na₂SO₄, filtered and evaporated in vacuo. Thematerial was treated with 5 mL warm EtOAc and the resulting solidfiltered off, rinsed with 1-2 mL additional EtOAc and dried to give thesemi-pure 4c as a tan-yellow powder (0.827 g). MS: m/z 347.1 (MH⁺).

C. 2-Chloro-N-(isoquinolin-5-yl)-2-(4-(trifluoromethyl)phenyl)acetamide.To a suspension of 4c (0.347 g, 1.00 mmol) in CHCl₃ (25 mL) was addedthionyl chloride (1 mL, 14 mmol). The reaction was stirred at ambienttemperature for 5 days, diluted with an additional 25 mL CHCl₃, washedtwice with 25 mL saturated NaHCO₃ then once with 25 mL brine. Theorganics were dried over Na₂SO₄ and filtered to give a solution ofcompound 4d in approximately 50 mL CHCl₃, about 0.02 M.

D.N-(Isoquinolin-5-yl)-2-(phenylthio)-2-(4-(trifluoromethyl)phenyl)acetamide.To the CHCl₃ solution of 4d (37-38 mL, 0.02 M, 0.75 mmol) was addedthiophenol (0.09 mL, 0.88 mmol) and an hour later more thiophenol (0.29mL, 2.84 mmol), CH₃CN (10 mL) and K₂CO₃ (0.211, 1.53 mmol). Afterstirring overnight the reaction mixture was diluted with 60 mL DCM andwashed with 50 mL 10% Na₂CO₃ solution then with 50 mL brine. Theorganics were dried with MgSO₄, filtered and evaporated in vacuo. Theresidue was purified by reverse-phase chromatography (25-95%acetonitrile/water+0.1% TFA). The proper fractions were frozen andlyophilized to give Compound 56 as a white powder (0.114 g). ¹H-NMR(DMSO-d₆): δ 10.66 (s, 1H), 9.50 (s, 1H), 8.56 (d, 1H), 8.10 (d, 1H),7.98 (d, 1H), 7.86 (d, 2H), 7.82-7.72 (m, 4H), 7.49 (d, 2H), 7.41-7.29(m, 3H), 5.76 (s, 1H); MS: m/z 439.1 (MH⁺).

Example 5

A. (4-Trifluoromethyl-phenyl)-acetic acid methyl ester. To a mixture ofCompound 3a (5.0 g, 24.5 mmol) in MeOH (25 mL) at room temperature wasadded conc. HCl (0.25 mL). The reaction was stirred at room temperaturefor 16 h. It was then concentrated under reduced pressure, diluted withCH₂Cl₂, washed sequentially with aq. 1N NaOH solution, H₂O and brine,dried over Na₂SO₄, and concentrated under reduced pressure to giveintermediate 5a (5.0 g, 94%).

B. 3-(2-Fluoro-phenyl)-2-(4-trifluoromethyl-phenyl)-propionic acidmethyl ester. To a solution of intermediate 5a (0.5 g, 2.29 mmol) in THF(25 mL) at −78° C. was added LDA (2.0 M in heptane/THF/ethylbenzene, 1.4mL, 2.8 mmol). The reaction mixture was stirred at −78° C. for 30 minbefore 2-fluorobenzyl bromide (0.42 ml, 3.48 mmol) was added. Thereaction was kept at −78° C. for 2 h, warmed to 0° C. and stirred foranother 2 h. To the reaction mixture was added aq. NH₄Cl solution, andthe resulting mixture was extracted with ethyl ether. The organicsolution was washed with brine, dried over Na₂SO₄, and concentratedunder reduced pressure. Purification by flash column chromatography(SiO₂, 10% EtOAc/heptane) afforded Compound 5b (0.49 g, 66%).

C. 3-(2-Fluoro-phenyl)-2-(4-trifluoromethyl-phenyl)-propionic acid. Amixture of Compound 5b (0.22 g, 0.67 mmol) and LiOH.H₂O (113 mg, 2.69mmol) in a mixed solution of THF (4 mL), MeOH (4 mL) and H₂O (4 mL) wasstirred at room temperature for 3 h. The reaction mixture was acidifiedwith aq. 10% HCl solution, and extracted with EtOAc. The organicsolution was washed with brine, dried over Na₂SO₄, and concentratedunder reduced pressure to afford Compound 5c (0.17 g, 81%).

D.3-(2-Fluoro-phenyl)-N-quinolin-5-yl-2-(4-trifluoromethyl-phenyl)-propionamide.To a solution of Compound 5c (51 mg, 0.16 mmol) and 5-aminoisoquinoline(47 mg, 0.33 mmol) in CH₂Cl₂ (3 mL) at room temperature was added Et₃N(0.14 mL, 1.0 mmol) followed by HATU (124 mg, 0.33 mmol). The reactionwas stirred at room temperature for 16 h. The reaction mixture was thendiluted with CH₂Cl₂, washed sequentially with aq. 10% HCl solution andaq. NaHCO₃ solution, dried over Na₂SO₄, and concentrated under reducedpressure. Purification by flash column chromatography (SiO₂, 40%EtOAc/heptane) afforded Compound 55 (30 mg, 42%). MS: 439 (M+1)⁺.

Example 6

A. 2-Hydroxy-N-quinolin-5-yl-2-(4-trifluoromethyl-phenyl)-acetamide. Toa solution of Compound 4a (1 mmol) and 5-aminoisoquinoline (1.5 mmol) inCH₂Cl₂ at room temperature may be added Et₃N (6 mmol) followed by HATU(1.5 mmol). The reaction may be stirred at room temperature for 16 h.The reaction mixture may then be diluted with CH₂Cl₂, washedsequentially with aq. NaHCO₃ solution and brine, dried over Na₂SO₄, andconcentrated under reduced pressure. Purification by flash columnchromatography may afford Compound 6a.

B.2-(2-Fluoro-phenoxy)-N-quinolin-5-yl-2-(4-trifluoromethyl-phenyl)-acetamide.To a solution of Compound 6a (1 mmol), 2-fluorophenol (2 mmol) and PPh₃(2 mmol) in THF (10 mL) at room temperature may be added DIAD (1.3mmol). The reaction may be stirred at room temperature. The mixture maythen be concentrated under reduced pressure, diluted with CH₂Cl₂, washedsequentially with aq. 1N NaOH solution and brine. The organic solutionmay be dried over Na₂SO₄ and concentrated under reduced pressure.Purification by flash column chromatography may afford Compound 6b.

Example 7

A. N-Indan-1-yl-2-phenylamino-2-(4-trifluoromethyl-phenyl)-acetamide,Cpd 10. To Compound 7a (0.15 g, 0.452 mmol) under argon was added(S)-1-aminoindane (0.060 g, 0.452 mmol), DMSO (1.5 mL), anddiisopropylethylamine (0.157 mL, 0.904 mmol). To this solution was addedHATU (0.21 g, 0.543 mmol) and the reaction mixture stirred for 16 hr atrt. The reaction mixture was diluted with acetonitrile to 2 mL, purifiedby gradient elution reverse-phase HPLC (CH₃CN, 0.1% TFA/H₂O, 0.1% TFA),and lyophilized to afford Compound 10. ¹H NMR (300 MHz, DMSO-d₆) δ ppm1.57-1.93 (m, 1 H), 2.24-2.48 (m, 1 H), 2.67-3.02 (m, 2 H), 5.12-5.34(m, 2 H), 6.50-7.36 (m, 9 H), 7.67-7.85 (m, 4 H), 8.78 (dd, J=17.71,8.29 Hz, 1 H); MS: C₂₄H₂₁F₃N₂O: m/z411.1 (M+1).

Example 8

A.2-(Indolin-1-yl)-N-(isoquinolin-5-yl)-2-(4-(trifluoromethyl)phenyl)acetamide.A solution of Compound 3b (2.298 g, 7.26 mmol) in DCM (40 mL) was addeddropwise over a period of 14 min to a solution of 5-amino isoquinoline(1.048 g, 7.27 mmol) in DCM (40 mL). The reaction mixture was stirred atroom temperature for an additional 30 min and then quenched withsaturated NaHCO₃ solution (40 mL). The organic phase was isolated, driedover Na₂SO₄ and diluted with DCM to a volume of approximately 80 mL.1/15^(th) of this solution by volume (0.48 mmol) was transferred to avial. To this was added indoline (0.11 mL, 0.98 mmol). After stirring atambient temperature for 2 days, the reaction mixture was washed oncewith saturated Na₂CO₃ solution (5 mL), dried over Na₂SO₄ and filtered.The organic phase was concentrated in vacuo and the residue was purifiedby reverse-phase chromatography (10-90% acetonitrile/water+0.1% TFA) toafford Compound 18 as a golden powder (0.103 g). ¹H-NMR (DMSO-d₆): δ10.73 (s, 1H), 9.59 (s, 1H), 8.61 (d, 1H), 8.19-8.12 (m, 2H), 8.02 (d,1H), 7.87-7.76 (m, 5H), 7.11-7.02 (m, 2H), 6.72 (d, 1H), 6.66 (t, 1H),5.80 (s, 1H), 3.74 (q, 1H), 3.20 (q, 1H), 3.02-2.84 (m, 2H); MS: m/z448.1 (MH⁺).

Following the procedure described above for Example 8 and substitutingthe appropriate reagents, starting materials and purification methodsknown to those skilled in the art, the following compounds of thepresent invention were prepared:

Cpd MS (M + 1)⁺ 19 462.1 26 462.1

Example 9

As a specific embodiment of an oral composition, 100 mg of Compound 1 isformulated with a sufficiently finely divided lactose to provide a totalamount of 580 to 590 mg to fill a size 0 hard gel capsule.

Biological Examples Example 1 In Vitro Canine TRPM8 Functional Assay

The functional activity of compounds of the formula (I) was determinedby measuring changes in intracellular calcium concentration using aCa²⁺-sensitive fluorescent dye. The changes in fluorescent signal weremonitored by a fluorescence plate reader, either a FLIPR™ (MolecularDevices) or FDSS (Hamamatsu). Increases in intracellular Ca²⁺concentration were readily detected upon activation with icilin.

HEK293 cells stably expressing canine TRPM8 were routinely grown asmonolayers in Dulbecco's minimum essential medium supplemented with 10%FBS, 2 mM L-glutamine, 100 units/mL penicillin, 100 ug/mL streptomycinand 400 μg/mL G418. Cells were maintained in 5% CO₂ at 37° C. At 24 hrsprior to assay, cells were seeded in black wall, clear-basepoly-D-lysine coated 384-well plates (BD Biosciences, NJ, USA) at adensity of 5,000 cells per well in culture medium and grown overnight in5% CO₂ at 37° C. On assay day, growth media was removed and cells wereloaded with Calcium 3 Dye (Molecular Devices) for 35 min at 37° C.,under 5% CO₂ and then for 25 min at room temperature and atmosphere.Subsequently, cells were tested for agonist-induced increases inintracellular Ca²⁺ levels using FLIPR™ or FDSS. Cells were challengedwith compounds of the formula (I) (at varying concentrations) andintracellular Ca²⁺ was measured for 5 min prior to the addition oficilin to all wells to achieve a final concentration that producesapproximately an 80% maximal response. EC₅₀ or IC₅₀ values for compoundsof the present invention were determined from eight-point dose-responsestudies and represent the concentration of compound required to elicitor inhibit 50% of the maximal response, respectively.

Maximal fluorescence intensity (FI) achieved upon addition of icilin wasexported from the FLIPR or FDSS software and further analyzed usingGraphPad Prism 3.02 (Graph Pad Software Inc., CA, U.S.A.). Basal FI wassubtracted prior to normalizing data to percent of maximal response.Curves were generated using the average of quadruplicate wells for eachdata point, were analyzed using nonlinear regression of either sigmoidaldose response or sigmoidal dose response (variable slope). Finally, theEC₅₀ and IC₅₀ values were calculated with the best-fit dose curvedetermined by GraphPad Prism. The resultant data are displayed in Table3.

TABLE 3 % Inh % Inh % Inh Cpd No. IC50 (μM) @ 1 μM @ 0.5 μM @ 0.2 μM 10.0358 102.92 2 59.612 3 0.301 101.11 4 0.072, 0.02885 102.46 104.04 50.1181 100 6 41.973 7 0.5287 100 8 0.4381 97.472 9 37.985 10 0.158 100.211 0.0891 103.62 12 0.2606 101.06 13 0.4888 86.882 14 30.961 15 27.04216 33.504 17 29.558 18 0.1031 103.95 19 0.5859 79.708 20 0.2327 101.1221 0.2866 95.294 22 0.0384 104.28 23 41.323 24 0.05485, 103.77 0.0442725 0.2734 99.008 26 39.484 27 0.0845 100.54 28 0.1609 98.259 29 0.441585.069 30 0.2573 96.049 31 0.017 102.16 32 37 33 38 34 0.226 84 35 0.07399 36 0.119 95 37 0.217 94 38 0.181 84 39 0.158 93 40 61 41 0.082 98 420.109 96 43 69 44 0.0968 98 45 52 46 72 47 23 48 0.094 98 49 0.092 83 500.1151 98.011 51 0.273 87 52 0.219 80 53 0.0653 90 54 0.1066 77 55 18 560.13

In Vivo Models Example 2 Inhibition of Icilin-Induced Behaviors inRodents

Icilin was initially developed as a “super-cooling” compound by DelmarChemicals Ltd. Subsequently it was shown to be one of the most potentknown agonists of TRPM8 (McKemy D D, et al., Nature 2002, 416(6876):52-8), having an EC₅₀=0.2 μM in stimulating calcium ion influx intoTRPM8 transfected cells (Behrendt H J, et al., Brit J Pharmacol 2004,141(4): 737-45). Initial in vivo testing of icilin showed it to cause“wet-dog” shakes in rats. Similar shaking or jumping behavior was alsoevident in mice, rabbits, cats, dogs and monkeys. In humans, icilinproduced a sensation of coolness on contact with mucous membranes, coldprickling when 0.1 mg was dropped on the tongue and coldness in themouth, pharynx and chest lasting 30-60 minutes when 5-10 mg was ingestedorally (Wei E T, Seid D A, J Pharm Pharmacol. 1983, 35, 110). Theinhibition or reversal of icilin-induced shaking behaviors in rodentsprovides evidence for the utility of TRPM8 antagonists of the formula(I) in modulating a disease or condition in a mammal in which thedisease or condition is affected by the modulation of TRPM8 receptors.

Example 2a Inhibition of Icilin-Induced “Wet-Dog” Shakes in Rats

Male Sprague Dawley rats (2200-450 g, Charles River Labs,n=6-9/treatment) were used to evaluate the ability of selected compoundsof the formula (I) to block icilin-induced “wet-dog” shakes (WDS).Compounds of the formula (I) were administered in an appropriatevehicle, such as, hydroxypropyl-β-cyclodextrin (HP β CD),methocellulose, 10% Solutol, or H₂O, or the like, by the appropriateroute, i.p. or p.o., 30-60 minutes before icilin. Icilin wasadministered in PEG-400 or 10% solutol/H₂O, at 1.0 or 3.0 mg/kg, i.p.and spontaneous “wet-dog” shakes were counted 10-20 minutes post-icilin.Results are presented as a percent inhibition of shakes, which wascalculated as [1−(test compound WDS count/vehicle WDS count)]×100.

Example 2b Reversal of Icilin-Induced Behaviors in Rats

Male Sprague Dawley rats (225-450 g, Charles River Labs,n=4-6/treatment) were used to evaluate the ability of selected compoundsof the formula (I) to reverse icilin-induced “wet-dog” shakes. Icilinwas administered in PEG-400 or 10% solutol/H₂O, at 1.0 or 3.0 mg/kg,i.p. and spontaneous “wet-dog” shakes (WDS) were counted 10-20 minutespost-icilin. Animals that exhibited 10 or more shakes were randomizedinto treatment groups and immediately administered compounds of theformula (I) in an appropriate vehicle, such as,hydroxypropyl-β-cyclodextrin (HP β CD), methocellulose, 10% Solutol, orH₂O, or the like, and by the appropriate route, such as, i.p. or p.o.Spontaneous “wet-dog” shakes were counted 60-70 minutes after compoundadministration. Results are presented as a percent inhibition of shakes,which was calculated as [1−(test compound WDS count/vehicle WDScount)]×100.

Example 3 In Vivo Model for of Chronic Inflammatory Pain: CompleteFreund's Adjuvant (CFA)-Induced Hyperalgesia

Intraplantar injection of complete Freund's adjuvant (CFA) in rodentsresults in a long-lasting inflammatory reaction, characterized by apronounced hypersensitivity to both thermal and mechanical stimuli. Thishypersensitivity peaks between 24-72 hours following injection and canlast for several weeks. To assess whether test compounds of the formula(I) reverse established hypersensitivity, a 100 μL intraplantarinjection of CFA (suspended in a 1:1 emulsion of saline and heat-killedMycobacterium tuberculosis in mineral oil) can be injected into a singlehind paw of Sprague-Dawley rats (typically males ranging from 150-350g). This paradigm also may be conducted with a multiple dosing or aprophylactic dosing regime designed to alter the course of hyperalgesiadevelopment. This test predicts the analgesic, anti-allodynic andantihyperalgesic effect of numerous effective clinical agents, includingacetaminophen, NSAIDS, such as, aspirin and ibuprofen, and opioids, suchas, morphine.

Example 3a CFA-Induced Paw Radiant Heat Hypersensitivity

Each rat is placed in a test chamber on a warm glass surface and allowedto acclimate for approximately 10 minutes. A radiant thermal stimulus(beam of light) is then focused through the glass onto the plantarsurface of each hind paw in turn. The thermal stimulus is automaticallyshut off by a photoelectric relay when the paw is moved or when thecut-off time is reached (20 seconds for radiant heat at ˜5 Amps). Aninitial (baseline) response latency to the thermal stimulus is recordedfor each animal prior to the injection of CFA. Twenty-four hoursfollowing intraplantar CFA injection, the response latency of the animalto the thermal stimulus is then re-evaluated and compared to theanimal's baseline response time. Only rats that exhibit at least a 25%reduction in response latency (i.e. hyperalgesia) are included infurther analysis. Immediately following the post-CFA latency assessment,test compound or vehicle (usually Solutol, hydroxypropylmethylcellulose, hydroxypropyl beta-cyclodextrin or PEG-400) isadministered i.p. or p.o. to rats. Post-compound treatment withdrawallatencies are assessed at fixed time intervals, typically 30, 60 and 120minutes. The percent reversal (% R) of hypersenstivitiy is calculatedaccording to the following formula:% Reversal=(Treatment Response−CFA Response)/(Baseline Response−CFAResponse)×100.

Cpd % Reversal No. Dose (mg/kg) Route @ 2 h 24 30 PO 28

Example 3b CFA-Induced Paw Cold Hypersensitivity

Prior to intraplantar CFA injection, mice or rats are placedindividually in elevated observation chambers having wire mesh floors.Through the mesh floor a series of three applications of acetone(0.04-0.10 mL/application) is sprayed onto the bottom of the paw using amultidose syringe device. A positive response takes the form of anabrupt withdrawal and licking of the paw. The cumulative duration oflicking is recorded for each of the three trials which are then averagedto give the individual's response. Twenty-four hours following CFAinjection acetone licking durations are markedly elevated implying ahypersensitivity to cooling. Test compounds of the formula (I) can beassessed for its ability to return acetone-evoked paw licking durationsto pre-CFA levels (typically near zero) following systemicadministration. Percent inhibition is calculated as follows% Inhibition=[1−(treatment licking duration/vehicle lickingduration)]×100.

Example 4 Chemically-Induced Abdominal Irritant Models of Visceral Pain

A chemical irritant (such as, acetic acid, kaolin, bradykinin,phenyl-p-(benzo)quinine, bromo-acetylcholine, or zymosan) is injected inmice intraperitoneally, causing a contraction of the abdominalmusculature, which is characterized by an elongation of the bodyextending through to the hind limbs. The number of such responses isquantitated and is reduced by pretreatment of analgesic agents, thusforming the basis for a screening test (Collier H O, et al., Br JPharmacol Chemother 1968, 32(2): 295-310). This type of abdominalirritant test has been used to predict the analgesic effect of numerousclinically effective agents, the potency of which in the abdominalirritant test parallels the magnitude of the dose needed in the reliefof clinical pain. Such agents include acetaminophen, NSAIDS, such as,aspirin and ibuprofen, opioids, such as, morphine and codeine, and othercentrally acting analgesics, such as, tramadol.

One modification of the chemically-induced abdominal irritant model ofvisceral pain is to pretreat animals with agents known to induceinflammatory responses following intraperitoneal injection (such as,LPS, zymosan, or thioglycolate). A small intraperitoneal dose of such aninflammogen, administered hours or days before the acute chemicalirritant challenge, has been shown to increase the number of abdominalcontractions observed (Ribeiro R A, et al., Eur J Pharmacol 2000,387(1): 111-8). While some analgesic agents are effective at mitigatingacute viscerochemical nociception, others, particularly those dependentupon receptor induction are more effective at preventing or reversingthe enhancement of behavioral responses caused by a preconditioninginflammatory stimulus. Because of the up-regulation of the TRPM8receptor in inflammation, TRPM8 antagonists that are effective atreducing the mean number of contractions are predicted to provideanalgesic action in human clinical use.

The ability of compounds of the formula (I) to mitigate chemicalirritant-induced abdominal contractions following a pre-conditioninginflammatory stimulus can be studied as follows. Thioglycolate (3%, w/v,2-3 mL i.p.) is injected into male CD1 mice (20-40 g, Charles RiverLabs), at a maximum dosage volume of 80 mL/kg, to induce peritonealinflammation. Following a twenty-four hour pre-inflammation period thesemice are dosed orally with compounds of the formula (I) (30 mg/kg; n=10)or vehicle (HPMC with 2% Tween80; n=9) and then one hour later subjectedto an abdominal irritant challenge of acetic acid (1%, 10 mL/kg, i.p.).Immediately following injection of acetic acid, mice are placedindividually in glass bell jars (approximately 15 cm in diameter) forcounting of abdominal contractions over the next 15 minutes. The totalnumber of abdominal contractions is summed for each treatment group andemployed in the following formula to calculate Percent Inhibition (% I):% I=[1−(test compound contractions/vehicle contractions)]×100.

Example 5 In Vivo Models of Neuropathic Pain

The sciatic nerve is the major sensorimotor innervation of the (hind)leg and foot. Injury to the sciatic nerve or its constituent spinalnerves often results in pain-related behaviors. In rats and mice, tightligation of the L5 spinal nerve with silk suture, partial tight ligationof the sciatic nerve with silk suture or loose ligation of the sciaticnerve with chromic gut suture each result in behaviors reminiscent ofneuropathic pain in humans. These lesions (one per animal) are performedsurgically in anesthetized rodents. Both the spinal nerve and sciaticnerve lesions result in allodynia, a painful response to normallyinnocuous stimuli, and hyperalgesia, an exaggerated response to normallynoxious stimuli. It is important to note that both of these pain-relatedbehaviors are evoked by the testing procedures and that normal use ofthe paw (e.g., walking) is relatively uncompromised, apart fromoccasional “guarding” of the paw. Subsequent to the surgery, thesubjects' behaviors, such as, grooming, feeding, and weight gain, arenormal, except for hypersensitivity (as defined above) of the affectedpaw.

In addition to induction by nerve damage resulting from accidentaltrauma or surgical procedures, neuropathic pain can also be induced bydiabetes (Fox, A, et al., Pain 1999, 81:307-316) or by treatment withchemotherapeutic agents, such as, paclitaxel or vincristine (Yaksh, T L,et al., Pain 2001, 93:69-76).

Agents that attenuate neuropathic pain in the clinic also are effectivein rodent neuropathic pain models. These agents include the recentlyapproved Cymbalta (Duloxetine, Iyengar, S., et al., JPET 2004311:576-584), morphine (Suzuki, R, et al., Pain 1999, 80:215-228) andgabapentin (Hunter, J C, et al., Eur J Pharmacol. 1997, 324:153-160).The dual TRPV1/TRPM8 receptor antagonist BCTC reduced mechanicalhyperalgesia and tactile allodynia in the chronic constriction injuryrodent neuropathic pain model (Pomonis, J D, et al., JPET. 2003,306:387-393; Behrendt, H, et al., Brit J Pharm. 2004, 141:737). Coldallodynia is a particularly debilitating symptom of neuropathic painconditions (Jorum E, et al. Pain 2003, 101:229-235). The antiallodyniceffect of compounds of the formula (I) in this rodent model ispredictive of clinical effect for these novel agents.

Example 5a Chronic Constriction Injury (CCl)-Induced Model ofNeuropathic Pain—Acetone-Induced Hypersensitivity

Male Sprague Dawley rats (225-450 g; n=5-8/treatment) were used toevaluate the ability of selected compounds of the formula (I) to reverseCCl-induced cold hypersensitivity. Four loose ligatures of 4-0 chromicgut were surgically placed around the left sciatic nerve underinhalation anesthesia as described by Bennett et al (Bennett G J, Xie YK, Pain 1988, 33(1): 87-107). Fourteen to 35 days following CCl surgery,subjects were placed in elevated observation chambers containing wiremesh floors and five applications of acetone (0.05 mL/applicationseparated by approximately 5 minutes) were spritzed onto the plantarsurface of the paw using a multidose syringe. An abrupt withdrawal orlifting of the paw was considered a positive response. The number ofpositive responses was recorded for each rat over the five trials.Following baseline withdrawal determinations, compounds of formula (I)are administered in an appropriate vehicle, such as,hydroxypropyl-β-cyclodextrin (HP β CD), methylcellulose, Methocel, 10%Solutol, or H₂O, or the like, by the appropriate route, i.p. or p.o. Thenumber of withdrawals were redetermined 1 to 3 h after compoundadministration. Results are presented as a percent inhibition of shakes,which was calculated for each subject as [1−(test compoundwithdrawals/pre-test withdrawals)]×100 and then averaged by treatment.

Example 5b Chronic Constriction Injury (CCl)-Induced Model ofNeuropathic Pain—Cold Plate-Induced Hypersensitivity

In male SD rats (175-325 g), four loose ligatures of 4-0 chromic gut aresurgically placed around the left sciatic nerve under inhalationanesthesia as described by Bennet et al. (Bennett G J, Xie Y K. Pain1988, 33(1):87-107). Seven to 21 days following sciatic chronicconstriction injury (CCl) surgery, the subjects can be placed onto acommercial cold plate device cooled by peltier elements such that thesurface temperature is held at 1° C. Each subject can undergo a 6 minuteconditioning period followed by a 3 minute assessment period duringwhich the total duration of hind paw lifting is recorded. This procedureis repeated at several intervals prior to and following systemic drugadministration. Compounds of the formula (I) can be assessed for theirability to return duration of paw lifting back to pre-lesion levels. Theduration of paw lifting during the 3 minute test period followingadministration of test compound is taken as a percentage of the durationof paw lifting during the 3 minute test period prior to test compoundtreatment.

Example 6 Inflammatory Agent-Induced Models of Pyresis/Antipyresis

Compounds of the formula (I) can be tested in animal models of pyresis,according to previously documented and validated methods, such as, thosedescribed by Kozak et al (Kozak W, Fraifeld V. Front Biosci 2004,9:3339-55). Fever is a frequent accompaniment of inflammatory disease.Animal models make use of the pyretic properties of yeast and otherinflammatory agents, injecting a yeast suspension or other agentsubcutaneously (Tomazetti J et al. J Neurosci Methods 2005,147(1):29-35); Van Miert A S, Van Duin C T., Eur J Pharmacol 1977, 44(3)197-204). For example, Male Wistar rats (75-100 g) can be housed ingroups of four to a cage at controlled temperature (23±1° C.) with a 12h light:12 h dark cycle (lights on at 07:00 h) and with standard labchow and tap water ad libitum. All measured temperatures can be takenbetween 08:00 and 19:00 h. Each animal can be used in only one study.Rectal temperature (TR) can be measured by inserting a lubricatedthermistor probe (external diameter: 3 mm) 2.8 cm into the rectum of theanimal. The probe can be linked to a digital device, which displayed thetemperature at the tip of the probe with a 0.1° C. precision and logsthe values over time. Immediately after measuring the initial basalrectal temperature, the animals can be injected with commerciallyavailable dried baker yeast (Saccharomyces cerevisiae) suspended inpyrogen-free 0.9% NaCl (0.05-0.25 g/kg, i.p.) or 0.9% NaCl (10 ml/kg).TR changes can be recorded every hour up to 12 h, and expressed as thedifference from the basal value. Since it has been previously reportedthat handling and temperature measuring-related stress alter rectaltemperature, these animals can be habituated to the injection andmeasuring procedure for 2 days before experiments are carried out. Inthese sessions, the animals can be subjected to the same temperaturemeasuring procedure described above, and can be injectedintraperitoneally (i.p.) with 0.9% NaCl (10 ml/kg).

To assess the effect of potential antipyretic compounds on basal rectaltemperature study animals can have their TR measured for 4 h, and afterthe fourth TR measurement they can be subcutaneously (s.c.) injectedwith vehicle (such as, 10% Solutol in sterile water 5 ml/kg) orcompounds of the formula (I) prepared in vehicle. TR can then berecorded every hour up to 8 h after the compound injections. To assessthe effect of compounds of the formula (I) on baker yeast-inducedhyperthermia, study animals can have their basal TR measured and then beinjected with a pyrogenic dose of baker yeast (for example, 0.135 g/kg).TR changes can be recorded every hour up to 4 h, when potentialantipyretics agents, such as, those compounds of the formula (I) areadministered. Rectal temperature can then be monitored over thefollowing 8 h. Basal rectal temperature and changes in rectaltemperature can be expressed as means±S.E.M. of the differences from TRat 07:00 h. Data can be analyzed by two-way analysis of variance(ANOVA), with time of measures treated as within subject factor,depending on the experimental design. Post hoc analysis can be carriedout by the F-test for simple effect and the Student-Newman-Keuls test,when appropriate. A value of P<0.05 would be considered statisticallysignificant.

The modification of the subsequent pyretic response by therapeuticagents can also be monitored by rectal telemetry or other measurementsof body temperature. Several clinically relevant agents, such as,acetaminophen, aspirin and ibuprofen, reduce fever in these models. Theantipyretic effect of TRPM8 antagonists, such as, compounds of theformula (I), in these tests would also be predictive of their clinicaleffect.

Example 7 CFA-Induced Model of Rheumatoid Arthritis

Compounds of the formula (I) can be tested in animal models ofrheumatoid arthritis, according to previously documented and validatedmethods, such as, those described by Nagakura et al (Nagakura Y, et al.,J Pharmacol Exp Ther 2003, 306(2): 490-7). For example, arthritis can beinduced by the CFA inoculation in the rats (Male Lewis rats 150-225 g;Charles River). Briefly, 100 mg of Mycobacterium butyricum (Difco,Detroit, Mich.) can be thoroughly mixed with 20 mL of paraffin oil. Thenmixture can be autoclaved for 20 min at 120° C. Each rat can be injectedin the right footpad (hind paw) with the mixture in a 0.1-mL volumeunder inhalation anesthesia. The rats serving as controls can beinjected with 0.1 mL of saline. Pain and other disease developmentparameters can be measured in the CFA- or saline-treated rats justbefore inoculation and up to 28 days post-inoculation. The measurementfor pain parameters can be conducted for both mechanical and thermal(hot or cold) endpoints. The measurement of mechanical allodynia can beperformed using the von Frey hairs (Semmes-Weinstein Monofilaments,Stoelting Co., IL) wherein the rats can be habituated to wire meshbottom cages before the start of the experiment. Static allodynia can betested in the unrestrained rats by touching the plantar surface of thehind paw with von Frey hairs in ascending order of force (1.2, 1.5, 2.0,3.6, 5.5, 8.5, 12, 15, 29, and 76 g) for up to 6 s or until a pawwithdrawal response can be elicited. The lowest amount of force requiredto elicit a response can be recorded as the withdrawal threshold in logg. Thermal hyperalgesia can be assessed using the radiant heat testwherein a mobile radiant heat source can be located under a glasssurface upon which the rat is placed. The beam of light can be focusedon the hind paw, and the paw withdrawal latencies are defined as thetime taken by the rat to remove its hind paw from the heat source. Themeasurement of joint hyperalgesia can be performed by a modification ofthe previously reported method (Rupniak N M J, et al., Pain. 1997,71:89-97). The torso of each rat can be held from the back with the leftpalm, and the bending and extension (one after the other and five timesin each direction) of the ankle within its limits of range of motion canbe performed with the right fingers. The total number of vocalizationsemitted after the manipulation (the bending and extension, five times ineach direction) can be recorded for each paw (the maximum score is 10for each paw).

The scoring of mobility can be performed by modifying the evaluationscale reported by Butler et al. (Butler S H, et al., Pain 1992,48:73-81): score 6, walks normally; score 5, walks being protectivetoward the ipsilateral hind paw (touches the ipsilateral hind paw fullyon the floor); score 4, walks being protective toward the ipsilateralhind paw (touches only the toe of the ipsilateral hind paw on thefloor); score 3, walks being protective toward both hind paws (touchesthe contralateral hind paw fully on the floor); score 2, walks beingprotective toward both hind paws (touches only the toe of thecontralateral hind paw on the floor); score 1, crawls only using thefore paws; and score 0, does not move. Paw volumes can be measured byvolume displacement of electrolyte solution in a commercially availableplethysmometer device. The hind paw can be immersed to the junction ofthe hairy skin, and the volumes can be read on a digital display. Thescoring of joint stiffness can be performed as follows: the body of ratscan be held from the back with the left palm, and the bending andextension (once in each direction) of the ankle within its limits ofrange of motion can be performed with the right fingers. It can beconfirmed beforehand that there is no restriction of ankle jointmovement in the bending and extension manipulations in naive rats, andthe scoring can be performed according to the evaluation scale reportedby Butler et al. (1992): score 2, there are restrictions of full rangeof movement of the ankle in both bending and extension; score 1, thereis a restriction of full range of movement of the ankle in bending orextension; and score 0, no restriction. The measurements for paw volumeand joint stiffness can be conducted for both hind paws.

Compounds of the formula (I) can be assessed for antihyperalgesicefficacy as follows: thirty-two rats (eight rats per dose and four dosesper compound) that are be treated with the CFA and another eight rats asnaive controls can be used for each drug evaluation. The analgesiceffects can be evaluated on post-inoculation day 9, when mechanicalallodynia, thermal hyperalgesia, joint hyperalgesia, and joint stiffnessin the ipsilateral paw reached almost the maximum, although thoseparameters in the contralateral paw changed only slightly and thesystemic disturbance shown by the change of mobility score is small. Onthe day before evaluation, body weight, mechanical allodynia, thermalhyperalgesia, and joint hyperalgesia can be measured for the 32 ratsthat are to be used for compound evaluation. The rats are allocated tofour groups (eight rats per group) such that the differences in theaverages of those parameters among the groups became small. All theanalgesic effect evaluations and behavioral observations can be carriedout by the observer who is blind to the drug treatment.

Data can be expressed as the mean+/−S.E.M. The time-course curves formechanical allodynia, thermal hyperalgesia, joint hyperalgesia, bodyweight, and paw volume can be subjected to two-way repeated measuresanalysis of variance with post hoc t test. In experiments for evaluationof compounds of formula (I), the difference in scores between thevehicle-treated and naive control groups can be analyzed by Student's ttest to confirm significant changes in the pain parameters in theipsilateral paw. The analgesic effects can be analyzed by Dunnett's ttest, and in each case the drug-treated groups can be compared with thevehicle-treated group. In each statistical analysis, the comparison canbe conducted for paws on the corresponding side. P<0.05 is consideredstatistically significant. In this model, the centrally actinganalgesics morphine and tramadol fully relieved pain, whereas theNSAIDs, indomethacin and diclofenac are partially effective, evidencingthe model's clinical predictability. The analgesic effect of compoundsof the formula (I) in this test would predict their clinical usefulnessin treating arthritis.

Example 8 In Vivo Model for Arthritis: Inflammogen-Induced Hyperalgesiaof the Knee Joint

Compounds of the formula (I) can be tested in animal models ofosteoarthritis, according to previously documented and validatedmethods, such as, those described by Sluka et al (Sluka K A, Westlund KN., Pain 1993, 55(3):367-77). For example, male Sprague-Dawley rats(Harlan, Indianapolis, Ind.) weighing 225 to 350 g can be brieflyanesthetized with vaporized halothane and then injected with a mixtureof 3% carrageenan and 3% kaolin (100 μL in 0.9% sterile saline) into thejoint cavity of one knee. After the injection, the animals can bereturned to their cages until the time of testing. For behavioraltesting animals can be placed in individual clear plastic cages on topof an elevated wire mesh surface that restricted movement. The animalsshould be allowed to acclimate for approximately 1 hour before testing.Von Frey filaments, as described above, can then be used to test forenhanced responses to mechanical stimuli. The filaments can besuccessively applied through the wire mesh perpendicularly to theplantar surface in between the pads of the third and fourth phalanges.The response threshold to mechanical stimuli can be determined beforeinflammation of the knee joint; 4 hours after inflammation to confirmthe development of hyperalgesia; immediately after the administration oftest compound, such as, those of Formula (I) i.e. 5 hours afterinflammation; and at 8, 12, and 24 hours after inflammation.

The Kruskal-Wallis test, a nonparametric test, can be used to analyzethe effects for frequency, intensity, and group for response tomechanical stimuli at baseline, 4 hours after inflammation, and aftercompound treatment (5 hours, 8 hours, 12 hours, and 24 hours afterinflammation). Further post hoc testing between groups can be executedby using the Mann-Whitney signed rank test. The data can be presented asmedian with 25th and 75th percentiles. Significance is P<0.05.

Additionally, the gait of the animal or other pain-related behavior canbe scored as the dependent measure of the painful effect of thearthritis on the animal's activity (Hallas B, Lehman S, Bosak A, et al.J Am Osteopath Assoc 1997, 97(4): 207-14). The effect of test drug onthe animal's normal behavior can be quantified from zero, meaning noresponse, to three for incapacitating impairment. Effective analgesictreatment includes the clinically used indomethacin (Motta A F, et al.,Life Sci 2003, 73(15):1995-2004). Thus the benefit of compounds of theformula (I) in this model would predict their clinical relevance.

Example 9 Sarcoma Cell-Induced Models of Bone Cancer Pain

Compounds of the formula (I) can be tested in animal models of bonecancer pain, according to previously documented and validated methods,such as those described in the scientific literature (El Mouedden M,Meert T F, Pharmacol Biochem Behav 2005, 82(1):109-19; Ghilardi J R, etal., J Neurosci 2005, 25(12):3126-31). In preparation for cellinoculation and tumor induction, osteolytic murine sarcoma cells (NCTC2472, American Type Culture Collection (ATCC), Rockville, Md., USA) canbe cultured in NCTC 135 medium (Invitrogen) containing 10% horse serum(Gibco) and passaged 2 times weekly according to ATCC guidelines. Fortheir administration, cells can be detached by scraping and thencentrifuged at 1000×g. The pellet can be suspended in fresh NCTC 135medium (2.5×10⁶ cells/20 μL) and then used for intramedullary femurinoculation. Male C3H/HeNCrI mice (25-30 g, Charles River Labs) can beused in such experiments. After induction of general anesthesia withxylazine (10 mg/kg i.p.) and ketamine (100 mg/kg i.p.) the left hind pawcan be shaved and disinfected with povidone-iodine followed by 70%ethanol. A superficial incision of 1 cm can then be made over the kneeoverlaying the patella. The patellar ligament can then be cut, exposingthe condyles of the distal femur. A 23-gauge needle can be inserted atthe level of the intercondylar notch and the intramedullary canal of thefemur to create a cavity for injection of the cells. Twenty microlitersof media (sham animals) or media containing tumor cells (approximately2.5×10⁶ cells) can then be injected into the bone cavity using asyringe. To prevent leakage of cells outside the bone, the injectionsite can be sealed with dental acrylic and the wound closed with skinstitches.

Pain behaviors can be evaluated in separate groups (n=6) of sham andbone tumor mice with confirmed hyperalgesia as assessed by spontaneouslifting behavior. Animals can be behaviorally tested during a 3-weekperiod prior to and after tumor inoculation. Body weight of the mice canbe recorded throughout the experimental period to help monitor generalhealth status. To measure the spontaneous lifting, the animals can behabituated in a transparent acrylic cylinder of 20 cm diameter put on anhorizontal surface and thereafter observed during 4 min for spontaneouslifting behavior of the left hind paw. After spontaneous liftingbehavior assessment, animals can be immediately placed on a mouserotarod (e.g. ENV-575M, Med Associates Inc., GA, USA) at a speed of 16rpm for 2 min wherein limb-use during forced ambulation is scored:4=normal; 3=limping; 2=partial non-use of left hind paw; 1=substantialnon-use of left hind paw; 0=non-use of left hind paw. Assessment of coldallodynia may be made by exposing the ipsilateral hind paw of the mouseto 5 repeated applications of acetone (20 μL) and quantifying thelift/licking frequency and/or duration. Post-mortem evaluation of bonedestruction can be assessed by ACT processing followed by scanning usinga system such as the Skyscan 1076\, microtomograph system for smallanimal imaging (Skyscan 1076\, Skyscan, Aartselaar, Belgium). Measuredhistomorphometry parameters of bone destruction can be subsequentlycorrelated with behavioral endpoints.

The antihyperalgesic, antiallodynic and disease modifying effects ofcompounds of the formula (I) can be tested in this murine model of bonecancer pain in separate groups (n=6 per dose group). Animals withconfirmed hyperalgesia, as assessed by spontaneous or acetone-evokedlifting, can be behaviorally tested, for example, on days 15 and 22after distal femur tumor inoculation before and 1 h after systemicadministration of vehicle (e.g. 10% Solutol in sterile water) orcompounds of the formula (I). The statistical analysis can be performedby one-way ANOVA to compare behavioral measurements and bone parametersamong the experimental groups. To compare behavioral measurements andbone parameters between sham and tumor-bearing animals, a Mann-Whitney Utest can be used. Results are considered statistically significant atP<0.05 (two-tailed). Data are expressed as mean±S.E.M.

Bone cancer causes intense pain in humans, mimicked in animal models ofbone cancer pain in rodents such as that described above. Analgesictreatments that are effective in this model include COX-2 inhibitors(Sabino M A, Ghilardi J R, Jongen J L, et al., Cancer Res 2002, 62(24):7343-9) and high doses of morphine (Luger N M et al., Pain 2002, 99(3):397-406), agents used clinically for pain relief in patientsexperiencing bone cancer pain. Because this model so closely mimics thehuman disease state, the finding that cold allodynia is a prominentsymptom (Lee, Seong et al., Yonsei Med J 2005, 46(2):252-9) stronglysupports the concept that TRPM8 antagonists of the present inventionwill provide relief of pain associated with human bone cancer.

Example 10 Respiratory Irritant-Induced Models of Cough

Compounds of the formula (I) can be tested in animal models ofantitussive activity, according to previously documented and validatedmethods, such as those described by: Tanaka, M. and Maruyama, K. J.Pharmacol. Sci. 2005, 99(1):77-82; Trevisani, M., et al., Throax 2004,59(9):769-72; and Hall, E., et al., J. Med. Microbiol. 1999, 48:95-98.Testing is conducted in transparent ventilated chambers with a constantairflow of 400 mL/min. The tussive agent (citric acid 0.25 M orcapsaicin 30 mM) can be nebulised via a miniultrasonic nebuliser with anoutput of 0.4 mL/min. The appearance of cough can be detected by meansof a tie clip microphone and confirmed by the characteristic posture ofthe animal. The cough sounds can be recorded and digitally stored. Ablinded observer subsequently counts the number of elicited coughefforts. In some cases, animals can be sensitized by pre-exposure tocertain agents, such as, ovalbumin. A test compound can be administeredto at the peak of irritant-induced cough to evaluate the antitussiveeffects of the compound. In addition, prophylactic or multiple dosingregimes can be utilized to evaluate the test compound for modulation ofthe onset and duration of irritant-induced cough. Variations of thesetests predict the antitussive effects of effective clinical agents,including NMDA antagonists, such as, dextrorphan and dextromethorphan,opioids, such as, codeine, beta 2 agonists, such as, salbutamol andantimuscarinics, such as, ipratropium (Bolser, D. C., et al., Eur. J.Pharmacol. 1995, 277(2-3):159-64; Braga, P. C., Drugs Exper. Clin. Res.1994, 20:199-203). The antitussive action of menthol in both guinea pigand humans Eccles R., Curr Allergy Asthma Rep 2003, 3(3):210-4; Laude EA, et al., Pulm Pharmacol. 1994, 7(3):179-84; Morice A H, et al., Thorax1994, 49(10):1024-6) is predictive of the clinical utility of compoundsof the formula (I) as antitussive agents.

Example 11 Chemical Irritant-Induced Models of Itch, Contact Dermatitis,Eczema and other Manifestations of Dermal Allergy, Hypersensitivityand/or Inflammation

Compounds of the formula (I) can be tested in animal models of contactdermatitis or itch, according to previously documented and validatedmethods, such as, those described in the scientific literature(Saint-Mezard P, et al., Eur J Dermatol. 2004, 14(5):284-95; Thomsen J.S., et al., J. Exp Dermatol. 2002, 11 (4):370-5; Weisshaar E, et al.,Arch Dermatol Res 1998, 290(6):306-11; Wille J J, et al., Skin PharmacolAppl Skin Physiol. 1999, 12(1-2):18-27). Mice (or species such as guineapig or rat) can be sensitized with 25 mL of 0.5% dinitrofluorobenzenesolution (DNFB diluted 4:1 in acetone:olive oil immediately beforeapplication or other haptens, such as, 12-myristate-13 acetate, picrylchloride, oxazolone, capsaicin, arachidonic acid, lactic acid,trans-retinoic acid or sodium lauryl sulfate) painted to the shaveddorsal skin or untreated (controls). Five days later, 10 mL of 0.2% DNFBa nonirritant dose) can be applied onto both sides of the right ear andthe same amount of solvent alone onto the left ear. Ear thickness can bemonitored daily using a caliper. Compounds of the formula (I) can beadministered at the peak of inflammation to evaluate the anti-allergyactivity of compounds. In addition, prophylactic or multiple dosingregimes can be utilized to evaluate the test compound for modulation ofthe onset and duration of anti-allergy activity. Variations of thesetests can predict the anti-allergy and itch activity of effectiveclinical agents. The ability of these models to predict the therapeuticeffect of compounds in human dermal conditions is supported by thecross-species ability of serotonin to induce itch (Weisshaar E, GollnickH., Skin Therapy Lett 2000, 5(5):1-2,5). Additionally, the contactsensitizing property of commercially important drugs and the ability ofion channel modulators to prevent and treat skin sensitization in thesemodels (Kydonieus A, et al., Proceedings of the International Symposiumon Controlled Release of Bioactive Materials 24th:23-24, 1997)demonstrate the therapeutic utility of compounds of the formula (I) indermal sensitization.

Example 12 Chemical Irritant-Induced Models of Rhinitis and otherManifestations of Nasal Hypersensitivity and/or Inflammation

Compounds of the formula (I) can be tested in animal models of rhinitis,according to previously documented and validated methods, such as thosedescribed in the scientific literature (Hirayama Y, et al., Eur JPharmacol. 2003, 467(1-3):197-203; Magyar T, et al., Vaccine 2002,20(13-14):1797-802; Tiniakov R L, et al., J Appl Physiol 2003,94(5):1821-8). Testing can be conducted in mouse, guinea pig, dog orhuman in response to intranasal challenge with one or more irritants,such as, cold air, capsaicin, bradykinin, histamine, pollens, dextransulfate, 2,4-tolylene diisocyanate, Bordetella bronchiseptica,Pasteurella multodica or acetic acid. In some cases, animals can besensitized by pre-exposure to certain agents including ragweed orovalbumin. Prior to or following irritant administration, the testsubject can receive, respectively, the prophylactic or therapeuticadministration one or more times of compounds of the formula (I), orvehicle control, by the enteral or parenteral route. Significantdifferences indicative of nasal rhinitis or sensitization for the testcompound-treated subjects compared with vehicle-treated subjects can betaken as evidence of anti-rhinitis activity. Independent variablesinclude dose, frequency and route of administration, time intervalbetween prophylactic or therapeutic test compound administration andirritant challenge as well as sex and non-sex genotype of the testsubject. The intimate role of neurogenic inflammation in thesehypersensitivity states demonstrates that compounds of the formula (I)desensitize or block the sensitization underlying these disease states.

Example 13 Conflict-Induced Models of Anxiety, Panic Disorder and otherNon-Adaptive Stressful or Phobic Responses

Compounds of the formula (I) can be tested in animal models of anxiety,panic disorders and other non-adaptive responses, according topreviously documented and validated methods, such as, those described byCryan and Holmes (Cryan J F, Holmes A., Nat Rev Drug Discov 2005,4(9):775-90) or Braw et.al. (Braw Y, et al., Behavioural Brain Research2006, 167:261-269). Specifically, for studies in rats, the followingapparati may be utilized: an open-field arena (62 cm×62 cm) enclosed byopaque walls (30 cm high) and plus-maze consists of two open arms, 50cm×10 cm, and two enclosed arms, 50 cm×10 cm×40 cm with an open roof,arranged such that the two arms of each type are opposite each other.The maze is elevated to a height of 70 cm. The walls of the enclosedarms are made from black Plexiglas, while the floors from whitePlexiglas. Videotape recordings can be analyzed using the ‘Observer’system (Noldus Information Technology). A subject rat can be removedfrom its home cage, weighed and placed gently in the center of theopen-field arena. The rat can be allowed to explore the open-fieldfreely while its behavior is videotaped for 5 min. Afterwards, it can betransferred to the plus-maze and placed at the center, facing a closedarm. The rat's behavior can again be videotaped for 5 min, after whichit can be returned to its home cage. The apparatus can cleaned using a70% ethanol solution between rats.

Open-field and plus-maze measures can be grouped into two behavioralclasses, namely ‘anxiety-like behaviors’ and ‘activity’. Open-fieldbehavioral measures may include 1) Anxiety measures: % time in centersquare, % number of entries to center square (from total squaresentered), % time freezing, latency to first freezing (freezing is scoredwhen the subject is in an immobile state for at least 3 seconds; and 2)Activity measures: Total squares entered, number of rearings (standingon two hind legs), latency for first rearing. Plus-maze measures mayinclude 1) Anxiety: % time in open arms, % number of entries to openarms (from total entries), number of unprotected head dips, latency toenter open arm; and 2) Activity: Total entries to all arms. Anxiety-likebehaviors and activity can be analyzed by one-way ANOVA's on each of themeasures, for each the between-subject comparisons. Plus-maze analysescan be conducted in a similar fashion.

Testing may also be conducted in mouse or rat in this fashion in orderto measure avoidance of other aversive environmental stimuli, such as,Geller or Vogel anticonflict tests, the light/dark test and thehole-board test (see Cryan J F, Holmes A., Nat Rev Drug Discov 2005,4(9):775-90). Prior to environmental exposure, the test subject canreceive the prophylactic administration one or more times of compoundsof the formula (I), or vehicle control (e.g. 10% Solutol in sterilewater), by the enteral or parenteral route. The cumulative time ornumber of times spent engaged in the aversive behavior can be measured.Significant differences in one or more of these measures for the testcompound-treated subjects compared with vehicle-treated subjects can betaken as evidence of anxiolytic activity. Because these models arepharmacologically validated by the effectiveness of clinically usefulanxiolytics (Cryan J F, Holmes A., Nat Rev Drug Discov 2005,4(9):775-90), they will be useful for the detection of anxiolyticcompounds of the formula (I).

Example 14 Bladder Pressure- and Hypertrophy-Induced Models of UrinaryIncontinence

Compounds of the formula (I) can be tested in animal models of urinaryincontinence according to previously documented and validated methods,such as those described by in the literature (Kaiser S, Plath T,(Metagen Pharmaceuticals GmbH, Germany). DE Patent Number 1,0215,321;McMurray G, et al., Br J Pharmacol 2006, 147 Suppl 2:S62-79). TRPM8 isexpressed in human prostate, testicle, seminiferous tubules, scrotalskin and inflamed bladder (Stein R J, et al., J Urol. 2004,172(3):1175-8); (Stein R J, et al., J Urol. 2004, 172(3):1175-8;Mukerji, et al., BMC Urology 2006, 6:6). Excitation of TRPM8 receptorsthrough cooling or application of menthol causes contraction in thebladder and a decrease in micturation threshold volume (Tsukimi Y,Mizuyachi K, et al., Urology. 2005, 65(2):406-10). To assess compoundsof the formula (I) for potential urinary incontinence activity,Sprague-Dawley rats are surgically implanted with bladder cathetersallowing for the delivery of fluid (typically saline) and the monitoringof pressure (using a pressure transducer). Cystometry recordings can bemonitored with a polygraph to evaluate voiding interval, thresholdpressure, bladder capacity, bladder compliance, and the number ofspontaneous bladder contractions. For example, the bladder catheter canbe connected to a Harvard infusion pump, and bladders perfused overnightwith saline at 2 mL/h. The next morning the bladder catheter can beattached (using a “T” connector) to a Statham pressure transducer (ModelP23Db) and to a Harvard infusion pump. A plastic beaker attached to aforce displacement transducer (Grass FTO3) can be placed under the rat'scage to collect and record urine volume. The cystometric evaluation ofbladder function can be started by infusing saline (20 mL/h) and afterthe first micturition the infusion is maintained for 20 min. Two hoursafter the first cystometry period, the rats can be dosed orally withcompounds of the formula (I) and a second cystometry is performedbetween 30 min and 4 h after administration of test compound. Theappropriate vehicle (e.g. 10% Solutol in sterile water) can be similarlyadministered to groups of rats that served as controls and thecystometry can be performed at the same respective time points.

Compounds of the formula (I) can also be evaluated under conditions ofbladder hypertrophy and instability. Under anesthesia, a silk ligatureis tied around the proximal urethra of rodents producing a partialoutlet obstruction and subsequent hypertrophied bladder developmentwithin 6-9 weeks (Woods M., et al., J. Urology. 2001, 166:1142-47).Cystometry recordings can then be evaluated as described above. Suchpreclinical procedures are sensitive to compounds having clinicalutility for the treatment of urinary incontinence (Soulard C, et al., JPharmacol Exp Ther 1992, 260(3):1152-8), and the activity of compoundsof the formula (I) in this model would be predictive of clinicalutility.

Example 15 In Vivo Model for Cold-Enhanced Central Pain States

Injury to the brain or spinal cord, such as that caused by trauma,interrupted blood flow or neurodegenerative diseases, often precipitatesa central pain condition. Examples of such injuries characterized, inpart by, a hypersensitivity to cold stimuli include multiple sclerosis(Morin C, et al., Clin J Pain 2002, 18(3):191-5; Svendsen K B, et al.,Pain 2005, 114(3):473-81), stroke or cerebral ischemia (Greenspan J D,et al., Pain. 2004, 109(3):357-66) and spinal cord injury (Defrin R,Ohry A, Blumen N, Urca G., Pain 2001, 89(2-3):253-63; Defrin R, et al.,Brain 2002, 125(Pt3):501-10; Finnerup N B, et al., Anesthesiology 2005,102(5):1023-30). Each of these conditions may be readily modeled inanimals for assessment of the ability of compounds of the formula (I) tomollify the hypersensitive state. For example, a spinal cord injury(SCI) can be performed in adult Sprague-Dawley rats having a body weightof 150-200 g at time of surgery (Erichsen, et al., Pain 2005,116:347-358). The rats can be anaesthetized with chloral hydrate (300mg/kg, i.p., Sigma, USA) and a catheter can be inserted into the jugularvein. A midline skin incision can then be made along the back to exposethe T11-L2 vertebrae. The animals can be positioned beneath a tunableargon ion laser (Innova model 70, Coherent Laser Products Division, CA,USA) operating at a wavelength of 514 nm with an average power of 0.17W.The laser light can be focused into a thin beam covering the single T13vertebra, which can be irradiated for 10 min. Immediately before theirradiation, erythrosin B (Aldrich, 32.5 mg/kg dissolved in 0.9% saline)can be injected intravenously via the jugular catheter. Due to rapidmetabolism of erythrosin B, the injection can be repeated after 5 min inorder to maintain adequate blood concentrations. During irradiation, thebody core temperature can be maintained at 37-38° C. by a heating pad.After irradiation the wound can be closed in layers and the skin suturedtogether.

SCI rats can be routinely tested for the presence of pain-like behaviorsfrom 3-4 weeks after surgery. The fur of the animals can be shaved atleast a day prior to examination of the cutaneous pain threshold toavoid sensitization of the skin receptors. During testing, the rats canbe gently held in a standing position by the experimenter and the flankarea and hindlimbs can be examined for hypersensitivity to sensorystimulation. On the day of drug testing, SCI rats can be administereddrug according to the experimental schedule and the time course ofpain-like behaviors can be measured. To test for the presence of coldallodynia, ethyl chloride or acetone can be sprayed onto the skin of theanimals, often that which has been previously determined to be sensitiveto mechanical stimulation by von Fry filament testing. The subsequentresponse to cold stimulation can be observed and classified according tothe following scale: 0, no visible response; 1, localized response (skintwitch) without vocalization; 2, transient vocalization; 3, sustainedvocalization. Kruskal Wallis ANOVA on ranks can be used to analyze theoverall effects of non-parametric data obtained in response to coldstimulation following pretreatment with either compounds of the formula(I) or vehicle.

Example 16 In Vivo Model for Post-Anesthetic Shivering

Spontaneous post-anesthetic tremor that resembles shivering is commonduring recovery from anesthesia. Risks to postoperative patients includean increase in metabolic rate of up to 400%, hypoxemia, wounddehiscence, dental damage, and disruption of delicate surgical repairs.The etiology of spontaneous post-anesthetic tremor is most commonlyattributed to normal thermoregulatory shivering in response tointraoperative hypothermia. In most operating and recovery rooms,shivering is controlled by the use of humidifiers, warming blankets, andinhalation of humidified heated oxygen. However, pharmacological controlis an effective alternate treatment modality (Bhatnagar S, et al.,Anaesth Intensive Care 2001, 29(2):149-54; Tsai Y C, Chu K S, AnesthAnalg 2001, 93(5):1288-92). Compounds of the formula (I) may be assessedfor their ability to mitigate post-ansethetic induced-shaking by usinganimal models such as that described by Nikki et al (Nikki P, TammistoT, Acta Anaesthesiol Scand 1968, 12(3): 125-34 and Grahn (Grahn, D A, etal., J Applied Physiology 1996, 81:2547-2554). For example, Wistar rats(males, weighing 250-450 g;) may be surgically implanted with an EEG/EMGrecording array to assess post anesthetic tremor activity. The EEGelectrodes are located bilaterally 2 mm off midline and adjacent tobregma and lamda. Following a one-week recovery period,frontal-occipital EEG, raw EMG, and integrated EMG activities, as wellas three temperatures (skin, rectal, and water blanket temperaturesduring anesthesia), and ambient temperature post-anesthesia can bemonitored throughout the experiment using copper-constantinthermocouples. The EEG and EMG signals can be recorded on polygraphpaper (5 mm/s, Grass model 7E polygraph) and, during recovery fromanesthesia, the EEG is computer scored in 10 second epochs as eithersynchronized: high amplitude (0.100 μV), low frequency (1-4 Hzdominated) activity characteristic of slow-wave sleep (SWS-like) ordesynchronized: low amplitude (75 μV), high frequency (5-15 Hzdominated), characteristic of waking and rapid-eye-movement sleep(W-like). The EMG activity can be quantified as the averaged summedvoltage/time interval by processing the raw EMG signal through anintegrator (Grass model 7P3, 0.5 s time constant). On the day of anexperiment, the animal can be placed in a small acrylic box (15×15×15cm) and exposed to a halothane vapor-air mixture (4% halothane).Immediately after the induction of anesthesia, the animal can be removedfrom the enclosure and subsequently anesthetized through a nose cone.Following cessation of anesthesia, two stages of recovery can be judged:emergence from anesthesia and restoration of behavioral activity(behavioral recovery). Emergence from anesthesia may be defined as anincrease in tonic EMG activity and a change in the EEG from a SWS-likepattern to a W-like pattern. Behaviorally, recovery has occurred whenthe animal rises from a prone position and initiated coordinatedmovements. The time intervals from termination of anesthesia toemergence and behavioral recovery can be measured in all animals. Timeinterval data can be subjected to a repeated measure analysis ofvariance, and the Scheffe's method can be employed for testingdifferences between pairs of means.

Example 17 TRPM8 Patch Clamp Assays

For patch clamp experiments, HEK293 cells are stably transfected withcanine TRPM8 and cultured in DMEM supplemented with 10% fetal bovineserum, 100 units/ml penicillin, 100 μg/ml streptomycin and 1 mg/ml G418.Cells are maintained at 37° C. and in 5% CO₂.

The extracellular solution contains (in mM): NaCl, 132; EGTA, 1; KCl,5.4; MgCl₂, 0.8; HEPES, 10; glucose, 10; pH=7.4. Recordings areperformed using the conventional whole-cell patch clamp technique, 1-2days after plating cells onto glass coverslips at densities appropriatefor single cell recording. Currents are amplified by a patch clampamplifier and filtered at 2 kHz (Axopatch 200B, Molecular Devices, UnionCity, Calif.). Menthol (100 μM) is applied to the cell at 0.5 ml/min viaa gravity-fed perfusion system. Recordings involving menthol activationare performed at 22° C.

In experiments where temperatures are varied, temperature ramps aregenerated by cooling the perfusate in an in-line cooler (Model SC-20,Warner Instruments, Hamden, Conn.) controlled by a temperaturecontroller (Model CL-100, Warner Instruments). The temperature in thevicinity of the recorded cell is measured with a custom-made miniaturethermo-microprobe connected to a monitoring thermometer (Model TH-8,Physitemp, Clifton, N.J.), and sampled using Digidata 1322A and pClamp9.0 (Molecular Devices), as are the currents concurrently measured inthe whole-cell patch clamp mode. The current is continuously sampled (at100 Hz) at a holding potential of −60 mV.

Compounds of the formula (I) are diluted from 10 mM DMSO stocks (storedat −20° C.) into an extracellular solution either containing 100 μMmenthol or subjected to cooling. Increasing concentrations of a compoundare applied to a cell in a cumulative manner and concentration-dependentresponses are measured after steady-state activation is achieved byeither 100 μM menthol or cooling to 10° C. A saturating concentration ofa reference antagonist is applied at the end of an experiment (either inthe presence of 100 μM menthol or 10° C. temperature) to establish thebaseline from which all the other measurements are subtracted.

Percentage inhibition by a compound is calculated as follows:100×(1−I_(comp)/I₀); where I_(comp) and I₀ are steady-state currentamplitudes in either the presence or absence of a concentration ofcompounds of the formula (I). Concentration-response data are fitted toa logistic function as follows: R=100/(1+c/IC₅₀)^(p); where, R is thepercentage inhibition, p is the Hill coefficient and c is theconcentration of compounds of the formula (I).

Example 18 In Vitro Rat and Human TRPM8 Functional Assay

For functional expression of TRPM8, the full-length cDNAs encoding humanand rat TRPM8 are subcloned into pCI-NEO mammalian expression vectors.The expression constructs are transiently transfected into HEK293 cellsaccording to the FuGENE 6 transfection reagent® (ROCHE) instructions.HEK293 cells are routinely grown as monolayers in Dulbecco's minimumessential medium supplemented with 10% FBS,1 mM L-glutamine, 100units/mL penicillin and 100 ug/mL streptomycin. Cells are maintained in5% CO₂ at 37° C. Within twenty-four hours, transiently transfected humanand rat TRPM8 are seeded into clear-base poly-D-lysine coated 384-wellplates (BD Biosciences, NJ, USA) at a density of 10,000 cells per wellin culture medium and grown overnight. The following day, all medium isremoved and the cells are incubated with 52 μL of 0.5×Calcium 3 Dye(Molecular Devices) prepared in complete assay buffer containing 20 mMHEPES, 0.1% BSA, and 2.5 mM probenecid at 37° C. for thirty fiveminutes. The cells are then incubated for an additional fifteen minutesat room temperature before initiating experiments. Following incubation,plates are inserted into a FDSS instrument, where cells were challengedwith compounds of the formula (I) (at varying concentrations) andintracellular Ca²⁺ are measured for 5 min prior to the addition of 100nM icilin. IC₅₀ values for compounds of compounds of the formula (I) aredetermined from eight-point dose-response studies

Maximal fluorescence intensity (FI) achieved upon addition of icilin isexported from the FDSS and further analyzed using GraphPad Prism 3.02(Graph Pad Software Inc., CA, U.S.A.). Basal FI is subtracted prior tonormalizing data to percent of maximal response. The dose responsecurves from the average of quadruplicate wells for each data point areanalyzed by using nonlinear regression of either sigmoidal dose responseor sigmoidal dose response (variable slope). Finally, the IC₅₀ valuesare calculated with the best-fit dose curve determined by Prism.

Example 19 Cold-Evoked Cardiovascular Pressor Responses

Compounds of the formula (I) can be tested in animals and humans fortheir ability to mitigate cardiovascular pressor responses evoked bycold exposure. Seasonal environmental cooling is directly associatedwith elevated blood pressure and an increased incidence of coronaryevents in human populations worldwide (Barnett, A G, et al., J EpidemiolCommunity Heath. 2005, 59 551-557). The clinical cold pressor testassesses changes in blood pressure (BP) and cold pain perception duringa 2-3 minute immersion of one hand into ice water. This test may beutilized to characterize analgesic compounds (Koltzenberg M, et al.,Pain 2006, 126(1-3):165-74) and to assess cold hypersensitivity(Desmeules J A, et al., Arthritis Rheum. 2003, 48(5):1420-9). Compoundsof the formula (I) can be studied in an anesthetized rat cold pressorparadigm to determine whether TRPM8 antagonism would interfere with theblood pressure pressor response to cold stimulation of the forepaws.Male Sprague-Dawley rats (300-450 g) anesthetized with sodiumpentobarbital are instrumented with a jugular catheter and an indwellingcarotid artery pressure transducer. Vehicle (10% Solutol in water) ortest compound is infused (1 mL/kg) over one minute through theintravenous catheter. Ten minutes later both forelimbs are packed incrushed ice for 5 minutes. Percent changes in mean arterial pressure inresponse to this cold stimulus are calculated for vehicle and testcompound pretreatments. Percent inhibition attributed to treatment withtest compound is then determined using the following formula: %Inhibition=[1−(cold evoked % change in BP post-test compound/cold evoked% change in BP post-vehicle)]×100.

While the foregoing specification teaches the principles of the presentinvention, with examples provided for the purpose of illustration, itwill be understood that the practice of the invention encompasses all ofthe usual variations, adaptations and/or modifications as come withinthe scope of the following claims and their equivalents.

We claim:
 1. A compound of Formula (I)

wherein A is phenyl; provided that R₂ is other than 2-chloro,2-C₁₋₄alkoxy, 4-C₁₋₄alkoxy, 2-C₁₋₄alkoxycarbonyl, or4-C₁₋₄alkoxycarbonyl; or L is —N(R₃)(CH₂)_(n)—, and R₁ and L areattached to adjacent carbon atoms of phenyl, R₁ and R₃ are optionallytaken with the atoms to which they are attached to formdihydro-indol-1-yl or dihydro-quinolin-1-yl; R₁ is one to threesubstituents selected from the group consisting of hydrogen, C₁₋₄alkyl,C₁₋₄alkoxy, fluoro, chloro, and hydroxy; L is —Z —(CH₂)_(n)— or —CH₂—; nis 0 or 1; Z is O, S, or NR₃; provided that when Z is O or S, A isphenyl; R₂ is hydrogen, C₁₋₄alkyl, C₁₋₄alkoxy, trifluoromethyl, chloro,fluoro, bromo, carboxy, or C₁₋₄alkoxycarbonyl; R₃ is hydrogen orC₁₋₃alkyl; R₄ is hydrogen or C₁₋₄alkyl; R₅ is isoquinolinyl andquinolinyl; wherein R₅ is optionally substituted with 1 to 2substituents independently selected from the group consisting ofC₁₋₄alkyl, hydroxy, C₁₋₄alkoxy, trifluoromethyl, chloro, fluoro, bromo,carboxy, C₁₋₄alkoxycarbonyl, and cyano; or an enantiomers, adiastereomer, or a pharmaceutically acceptable salt thereof; providedthat a compound of Formula (I) is other than a compound wherein A isphenyl, R₁ is 3-chloro, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, nis 1, Z is NR₃, R₃ is hydrogen, R₄ is hydrogen, and R₅ isisoquinolin-5-yl; and a compound wherein A is phenyl, R₁ is 4-methoxy,R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ is hydrogen, R₄is hydrogen, and R₅ is isoquinolin-5-yl.
 2. The compound according toclaim 1 wherein R₂ is other than 2-chloro, 2-C₁₋₄alkoxy, 4-C₁₋₄alkoxy,2-C₁₋₄alkoxycarbonyl, or 4-C₁₋₄alkoxycarbonyl; or L is —N(R₃)(CH₂)_(n)—,and R₁ and L are attached to adjacent carbon atoms of phenyl, R₁ and R₃are optionally taken with the atoms to which they are attached to formdihydro-indol-1-yl or dihydro-quinolin-1-yl.
 3. The compound accordingto claim 2 wherein R₂ is other than 2-chloro, 2-C₁₋₄alkoxy,4-C₁₋₄alkoxy, 2-C₁₋₄alkoxycarbonyl, or 4-C₁₋₄alkoxycarbonyl; and, when Lis —N(R₃)(CH₂)_(n)—, and R₁ and L are attached to adjacent carbon atomsof phenyl, R₁ and R₃ are optionally taken with the atoms to which theyare attached to form dihydro-indol-1-yl.
 4. The compound according toclaim 3 wherein R₂ is other than 2-chloro, 2-C₁₋₄alkoxy, 4-C₁₋₄alkoxy,2-C₁₋₄alkoxycarbonyl, or 4-C₁₋₄alkoxycarbonyl.
 5. The compound accordingto claim 1 wherein R₁ is one to two substituents selected from the groupconsisting of hydrogen, C₁₋₄alkyl, C₁₋₄alkoxy, fluoro, chloro, andhydroxy.
 6. The compound according to claim 5 wherein R₁ is one to twosubstituents selected from the group consisting of hydrogen, C₁₋₃alkyl,C₁₋₃alkoxy, fluoro, and chloro.
 7. The compound according to claim 6wherein R₁ is one to two substituents selected from the group consistingof hydrogen, methyl, fluoro, and chloro.
 8. The compound according toclaim 1 wherein L is —Z—(CH₂)_(n)— or —CH₂—; wherein n is 0 or 1; and Zis S or NR₃; provided that Z is S.
 9. The compound according to claim 8wherein L is —Z—(CH₂)_(n)—; wherein n is 0; and Z is S or NR₃; providedthat Z is S.
 10. The compound according to claim 9 wherein L is—Z—(CH₂)_(n)—; wherein n is 0; and Z is NR₃.
 11. The compound accordingto claim 1 wherein R₂ is hydrogen, C₁₋₄alkyl, trifluoromethyl, chloro,or bromo; provided that R₂ is other than 2-chloro.
 12. The compoundaccording to claim 1 wherein R₂ is t-butyl, trifluoromethyl, chloro, orbromo; provided that R₂ is other than 2-chloro.
 13. The compoundaccording to claim 1 wherein is 4-t-butyl, 4-trifluoromethyl, or4-chloro.
 14. The compound according to claim 1 wherein R₃ is hydrogenor methyl.
 15. The compound according to claim 1 wherein R₃ is hydrogen.16. The compound according to claim 1 wherein R₄ is hydrogen.
 17. Thecompound according to claim 1 wherein R₅ quinolinyl or isoquinolinyl;wherein R₅ is optionally substituted with one to two substituentsindependently selected from the group consisting of C₁₋₄alkyl,C₁₋₄alkoxy, and fluoro.
 18. The compound according to claim 17 whereinR₅ is quinolinyl or isoquinolinyl and is optionally substituted with oneC₁₋₄alkyl substituent.
 19. The compound according to claim 1 wherein R₅is quinolin-5-yl or isoquinolin 5yl.
 20. A compound of Formula (I)

wherein A is phenyl; provided that R₂ is other than 2-chloro; or, L is—N(R₃)(CH₂)_(n)—, and R₁ and L are attached to adjacent carbon atoms, R₁and R₃ are optionally taken with the atoms to which they are attached toform dihydro-indol-1-yl or dihydro-quinolin-1-yl; R₁ is one to twosubstituents selected from the group consisting of hydrogen, C₁₋₄alkyl,C₁₋₄alkoxy, fluoro, chloro, and hydroxl; L is —Z—(CH₂)_(n)— or —CH₂—;wherein n is 0 or 1; and Z is S or NR₃; provided that when Z is S, A isphenyl R₂ is hydrogen, C₁₋₄alkyl, trifluoromethyl, chloro, or bromo; R₃is hydrogen or methyl; R₄ is hydrogen; and R₅ is quinolinyl orisoquinolinyl, wherein R₅ is optionally substituted with 1-2substituents independently selected from the group consisting ofC₁₋₄alkyl, C₁₋₄alkoxy, and fluoro; or an enantiomers, a diastereomer, ora pharmaceutically acceptable salt thereof; provided that a compound ofFormula (I) is other than a compound wherein R₁ is 3-chloro, R₂ is4-trifluoromethyl, L is —Z—(CH₂)_(n)—, n is 1, Z is NR₃, R₃ is hydrogen,R₄ is hydrogen, and R₅ is isoquinolin-5-yl or a compound wherein R₁ is4-methoxy, R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ ishydrogen, R₄ is hydrogen, and R₅ is isoquinolin-5-yl.
 21. A compound ofFormula (I)

wherein A is phenyl; provided that R₂ is other than 2-chloro; or, L is—N(R₃)(CH₂)_(n)—, and R₁ and L are attached to adjacent carbon atoms, R₁and R₃ are optionally taken with the atoms to which they are attached toform dihydro-indol-1-yl; R₁ is one to two substituents selected from thegroup consisting of hydrogen, C₁₋₃alkyl, C₁₋₃alkoxy, fluoro, and chloro;L is —Z—(CH₂)_(n)—; wherein n is 0; and Z is S or NR₃; R₂ is t-butyl,trifluoromethyl, chloro, or bromo; R₃ is hydrogen; R₄ is hydrogen; andR₅ is quinolinyl or isoquinolinyl wherein R₅ is optionally substitutedwith one to two C₁₋₄alkyl substituents; or an enantiomers, adiastereomer, or a pharmaceutically acceptable salt thereof; providedthat a compound of Formula (I) is other than a compound wherein R₁ is4-methoxy, R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, n is 0, Z is NR₃, R₃ ishydrogen, R₄ is hydrogen, and R₅ is isoquinolin-5-yl.
 22. A compound ofFormula (I)

wherein A is phenyl; provided that R₂ is other than 2-chloro; R₁ is oneto two substituents selected from the group consisting of hydrogen,C₁₋₃alkyl, methoxy, fluoro, and chloro; L is —Z—(CH₂)_(n)—; wherein n is0; and Z is NR₃; R₂ is t-butyl, trifluoromethyl, chloro, or bromo; R₃and R₄ are each hydrogen; and R₅ is quinolinyl or isoquinolinyl; or anenantiomers, a diastereomer, or a pharmaceutically acceptable saltthereof; provided that a compound of Formula (I) is other than acompound wherein R₁ is 4-methoxy, R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, nis 0, Z is NR₃, R₃ is hydrogen, R₄ is hydrogen, and R₅ isisoquinolin-5-yl.
 23. A compound of Formula (I)

wherein A is phenyl; R₁ is one to two substituents selected from thegroup consisting of hydrogen, methyl, fluoro, and chloro; L is—Z—(CH₂)_(n)—; wherein n is 0; and Z is NR₃; R₂ is 4-t-butyl,4-trifluoromethyl, or 4-chloro; R₃ and R₄ are each hydrogen; and R₅ isquinolin-5-yl or isoquinolin-5-yl; or an enantiomers, a diastereomer, ora pharmaceutically acceptable salt thereof.
 24. A compound of Formula(I)

Formula (I)selected from the group consisting of a compound of Formula(I) wherein A is phenyl, R₁ is H, R₂ is 4-t-butyl, L is —Z—(CH₂)_(n)—, Zis NR₃, R₃ is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compoundof Formula (I) wherein A is phenyl, R₁ is t-butyl, R₂ is H, L is—Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and R₅ isisoquinolin-5-yl; a compound of Formula (I) wherein A is phenyl, R₁ isH, R₂ is 4-t-butyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is methyl, n is 0,R₄ is H, and R₅ is isoquinolin-5-yl; a compound of Formula (I) wherein Ais phenyl, R₁ is H, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z isNR₃, R₃ is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compound ofFormula (I) wherein A is phenyl, R₁ is H, R₂ is 4-t-butyl, L is—Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and R₅ isquinolin-6-yl; a compound of Formula (I) wherein A is phenyl, R₁ is H,R₂ is 4-t-butyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H,and R₅ is isoquinolin-6-yl; a compound of Formula (I) wherein A isphenyl, R₁ is 2-chloro, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Zis NR₃, R₃ is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compoundof Formula (I) wherein A is phenyl, R₁ is 3-chloro, R₂ is4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ isH, and R₅ is isoquinolin-5-yl; a compound of Formula (I) wherein A isphenyl, R₁ is 4-chloro, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Zis NR₃, R₃ is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compoundof Formula (I) wherein A is phenyl, R₁ is 2-chloro, R₂ is4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 1, R₄ isH, and R₅ is isoquinolin-5-yl; a compound of Formula (I) wherein A isphenyl, R₁ is 4-chloro, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Zis NR₃, R₃ is H, n is 1, R₄ is H, and R₅ is isoquinolin-5-yl; a compoundof Formula (I) wherein A is phenyl, R₁ is taken with R₃ to form2,3-dihydro-indol-1-yl, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Zis NR₃, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compound ofFormula (I) wherein A is phenyl, R₁ is taken with R₃ to form3,4-dihydro-2H-quinolin-1-yl, R₂ is 4-trifluoromethyl, L is—Z—(CH₂)_(n)—, Z is NR₃, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; acompound of Formula (I) wherein A is phenyl, R₁ is 2-methoxy, R₂ is4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ isH, and R₅ is isoquinolin-5-yl; a compound of Formula (I) wherein A isphenyl, R₁ is 2-methyl, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Zis NR₃, R₃ is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compoundof Formula (I) wherein A is phenyl, R₁ is 2-isopropyl, R₂ is4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ isH, and R₅ is isoquinolin-5-yl; a compound of Formula (I) wherein A isphenyl, R₁ is 2-fluoro, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Zis NR₃, R₃ is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compoundof Formula (I) wherein A is phenyl, R₁ is 2,6-dimethyl, R₂ is4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ isH, and R₅ is isoquinolin-5-yl; a compound of Formula (I) wherein A isphenyl, R₁ is taken with R₃ to form 7-methyl-2,3-dihydro-indol-1-yl, R₂is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, n is 0, R₄ is H, andR₅ is isoquinolin-5-yl; a compound of Formula (I) wherein A is phenyl,R₁ is H, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H,n is 0, R₄ is H, and R₅ is quinolin-8-yl; a compound of Formula (I)wherein A is phenyl, R₁ is H, R₂ is 4-trifluoromethyl, L is—Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and R₅ isisoquinolin-5-yl (*S); a compound of Formula (I) wherein A is phenyl, R₁is H, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, nis 0, R₄ is H, and R₅ is isoquinolin-5-yl (*R); a compound of Formula(I) wherein A is phenyl, R₁ is H, R₂ is 4-bromo, L is —Z—(CH₂)_(n)—, Zis NR₃, R₃ is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compoundof Formula (I) wherein A is phenyl, R₁ is 4-fluoro, R₂ is4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ isH, and R₅ is isoquinolin-5-yl; a compound of Formula (I) wherein A isphenyl, R₁ is 3-fluoro, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Zis NR₃, R₃ is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compoundof Formula (I) wherein A is phenyl, R₁ is H, R₂ is 4-chloro, L is—Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and R₅ isisoquinolin-5-yl; a compound of Formula (I) wherein A is phenyl, R₁ is2-fluoro, R₂ is 4-bromo, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0,R₄ is H, and R₅ is isoquinolin-5-yl; a compound of Formula (I) wherein Ais phenyl, R₁ is H, R₂ is 3-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z isNR₃, R₃ is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compound ofFormula (I) wherein A is phenyl, R₁ is 2-fluoro, R₂ is 4-chloro, L is—Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and R₅ isisoquinolin-5-yl; a compound of Formula (I) wherein A is phenyl, R₁ is4-fluoro, R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0,R₄ is H, and R₅ is isoquinolin-5-yl; a compound of Formula (I) wherein Ais phenyl, R₁ is 2,4-difluoro, R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z isNR₃, R₃ is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compound ofFormula (I) wherein A is phenyl, R₁ is 4-methoxy, R₂ is 4-chloro, L is—Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and R₅ isquinolin-5-yl; a compound of Formula (I) wherein A is phenyl, R₁ is2,6-difluoro, R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, nis 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compound of Formula (I)wherein A is phenyl, R₁ is 2,6-difluoro, R₂ is 4-chloro, L is—Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and R₅ isquinolin-5-yl; a compound of Formula (I) wherein A is phenyl, R₁ is2,4-difluoro, R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃is H, n is 0, R₄ is H, and R₅ is isoquinolin-5-yl; a compound of Formula(I) wherein A is phenyl, R₁is 2,3-difluoro, R₂ is 4-chloro, L is—Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and R₅ isisoquinolin-5-yl; a compound of Formula (I) wherein A is phenyl, R₁is2,3-difluoro, R₂ is 4-chloro, L is —Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, nis 0, R₄ is H, and R₅ is quinolin-5-yl; a compound of Formula (I)wherein A is phenyl, R₁is 2,5-difluoro, R₂ is 4-chloro, L is—Z—(CH₂)_(n)—, Z is NR₃, R₃ is H, n is 0, R₄ is H, and R₅ isquinolin-5-yl; a compound of Formula (I) wherein A is phenyl, R₁is2-fluoro, R₂ is 4-trifluoromethyl, L is —-CH₂—, R₄ is H, and R₅ isisoquinolin-5-yl; a compound of Formula (I) wherein A is phenyl, R₁is H,R₂ is 4-trifluoromethyl, L is —Z—(CH₂)_(n)—, Z is S, R₃ is H, n is 0, R₄is H, and R₅ is isoquinolin-5-yl; or an enantiomers, a diastereomer, ora pharmaceutically acceptable salt thereof.
 25. A pharmaceuticalcomposition comprising a compound of claim 1 and at least one of apharmaceutically acceptable carrier, a pharmaceutically acceptableexcipient, and a pharmaceutically acceptable diluent.
 26. Thepharmaceutical composition of claim 25, wherein the composition is asolid oral dosage form.
 27. A pharmaceutical composition of claim 25,wherein the composition is a syrup, an elixir, or a suspension.